Compounds and methods for inhibiting hepatitis C viral replication

ABSTRACT

Macrocyclic compounds having the structures described herein are useful for inhibiting replication of the hepatitis C virus (HCV). In preferred embodiments, the compounds are active against both the NS3 protease and the NS3 helicase of HCV.

RELATED APPLICATION

This Application is the U.S. National Phase under 35 U.S.C. §371 ofInternational Application No. PCT/US2006/040049, filed Oct. 10, 2006(published as WO 07/044,893 and herein incorporated by reference), whichclaims the benefit of U.S. Provisional Application No. 60/725,564, filedOct. 11, 2005, which is incorporated herein by reference in itsentirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to compounds, processes for theirsynthesis, compositions and methods for the treatment of hepatitis Cvirus (HCV) infection.

2. Description of the Related Art

Hepatitis C virus (HCV) infection is the most common chronic blood borneinfection in the United States. Although the numbers of new infectionshave declined, the burden of chronic infection is substantial, withCenters for Disease Control estimates of 3.9 million (1.8%) infectedpersons in the United States. Chronic liver disease is the tenth leadingcause of death among adults in the United States, and accounts forapproximately 25,000 deaths annually, or approximately 1% of all deaths.Studies indicate that 40% of chronic liver disease is HCV-related,resulting in an estimated 8,000-10,000 deaths each year. HCV-associatedend-stage liver disease is the most frequent indication for livertransplantation among adults.

Antiviral therapy of chronic hepatitis C has evolved rapidly over thelast decade, with significant improvements seen in the efficacy oftreatment. Nevertheless, even with combination therapy using pegylatedIFN-α plus ribavirin, 40% to 50% of patients fail therapy, i.e., arenonresponders or relapsers. These patients currently have no effectivetherapeutic alternative. In particular, patients who have advancedfibrosis or cirrhosis on liver biopsy are at significant risk ofdeveloping complications of advanced liver disease, including ascites,jaundice, variceal bleeding, encephalopathy, and progressive liverfailure, as well as a markedly increased risk of hepatocellularcarcinoma.

The high prevalence of chronic HCV infection has important public healthimplications for the future burden of chronic liver disease in theUnited States. Data derived from the National Health and NutritionExamination Survey (NHANES III) indicate that a large increase in therate of new HCV infections occurred from the late 1960s to the early1980s, particularly among persons between 20 to 40 years of age. It isestimated that the number of persons with long-standing HCV infection of20 years or longer could more than quadruple from 1990 to 2015, from750,000 to over 3 million. The proportional increase in persons infectedfor 30 or 40 years would be even greater. Since the risk of HCV-relatedchronic liver disease is related to the duration of infection, with therisk of cirrhosis progressively increasing for persons infected forlonger than 20 years, this will result in a substantial increase incirrhosis-related morbidity and mortality among patients infectedbetween the years of 1965-1985.

HCV is an enveloped positive strand RNA virus in the Flaviviridaefamily. The single strand HCV RNA genome is approximately 9500nucleotides in length and has a single open reading frame (ORF) encodinga single large polyprotein of about 3000 amino acids. In infected cells,this polyprotein is cleaved at multiple sites by cellular and viralproteases to produce the structural and non-structural (NS) proteins ofthe virus. In the case of HCV, the generation of mature nonstructuralproteins (NS2, NS3, NS4, NS4A, NS4B, NS5A, and NS5B) is effected by twoviral proteases. The first viral protease cleaves at the NS2-NS3junction of the polyprotein. The second viral protease is serineprotease contained within the N-terminal region of NS3 (herein referredto as “NS3 protease”). NS3 protease mediates all of the subsequentcleavage events at sites downstream relative to the position of NS3 inthe polyprotein (i.e., sites located between the C-terminus of NS3 andthe C-terminus of the polyprotein). NS3 protease exhibits activity bothin cis, at the NS3-NS4 cleavage site, and in trans, for the remainingNS4A-NS4B, NS4B-NS5A, and NS5A-NS5B sites. The NS4A protein is believedto serve multiple functions, acting as a cofactor for the NS3 proteaseand possibly assisting in the membrane localization of NS3 and otherviral replicase components. Apparently, the formation of the complexbetween NS3 and NS4A is necessary for NS3-mediated processing events andenhances proteolytic efficiency at all sites recognized by NS3. The NS3protease also exhibits nucleoside triphosphatase and RNA helicaseactivities. NS5B is an RNA-dependent RNA polymerase involved in thereplication of HCV RNA.

LITERATURE

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SUMMARY OF THE INVENTION

A first embodiment provides a compound selected from the groupconsisting of:

A second embodiment provides a pharmaceutical composition, comprising apharmaceutically acceptable excipient; and a pharmaceutically effectiveamount of the compound recited in the first embodiment.

A third embodiment provides a method of modulating the activity of ahepatitis C virus, comprising contacting the virus with an effectiveamount of the compound of the first embodiment or the pharmaceuticalcomposition of the second embodiment.

A fourth embodiment provides a method of treating a mammal, comprising:identifying a mammal infected with a hepatitis C virus; andadministering an effective amount of the compound of the firstembodiment or the pharmaceutical composition of the second embodiment tothe mammal.

A fifth embodiment provides a method of modulating the activity of ahepatitis C virus, comprising:

selecting a compound having both NS3 protease and NS3 helicase activity,wherein the compound is selected from the group consisting of

contacting the compound with a hepatitis C virus in an amount effectiveto modulate both NS3 protease activity and NS3 helicase activity.

A sixth embodiment provides a method of treating a mammal, comprising:

identifying a mammal infected with a hepatitis C virus;

identifying a compound having both NS3 protease and NS3 helicaseactivity, wherein the compound is selected from the group consisting of

administering an effective amount of the compound to the mammal.

These and other embodiments are described in greater detail below.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Definitions

As used herein, the term “hepatic fibrosis,” used interchangeably hereinwith “liver fibrosis,” refers to the growth of scar tissue in the liverthat can occur in the context of a chronic hepatitis infection.

The terms “individual,” “host,” “subject,” and “patient” are usedinterchangeably herein, and refer to a mammal, including, but notlimited to, primates, including simians and humans.

As used herein, the term “liver function” refers to a normal function ofthe liver, including, but not limited to, a synthetic function,including, but not limited to, synthesis of proteins such as serumproteins (e.g., albumin, clotting factors, alkaline phosphatase,aminotransferases (e.g., alanine transaminase, aspartate transaminase),5′-nucleosidase, γ-glutaminyltranspeptidase, etc.), synthesis ofbilirubin, synthesis of cholesterol, and synthesis of bile acids; aliver metabolic function, including, but not limited to, carbohydratemetabolism, amino acid and ammonia metabolism, hormone metabolism, andlipid metabolism; detoxification of exogenous drugs; a hemodynamicfunction, including splanchnic and portal hemodynamics; and the like.

The term “sustained viral response” (SVR; also referred to as a“sustained response” or a “durable response”), as used herein, refers tothe response of an individual to a treatment regimen for HCV infection,in terms of serum HCV titer. Generally, a “sustained viral response”refers to no detectable HCV RNA (e.g., less than about 500, less thanabout 200, or less than about 100 genome copies per milliliter serum)found in the patient's serum for a period of at least about one month,at least about two months, at least about three months, at least aboutfour months, at least about five months, or at least about six monthsfollowing cessation of treatment.

“Treatment failure patients” as used herein generally refers toHCV-infected patients who failed to respond to previous therapy for HCV(referred to as “non-responders”) or who initially responded to previoustherapy, but in whom the therapeutic response was not maintained(referred to as “relapsers”). The previous therapy generally can includetreatment with IFN-α monotherapy or IFN-α combination therapy, where thecombination therapy may include administration of IFN-α and an antiviralagent such as ribavirin.

As used herein, the terms “treatment,” “treating,” and the like, referto obtaining a desired pharmacologic and/or physiologic effect. Theeffect may be prophylactic in terms of completely or partiallypreventing a disease or symptom thereof and/or may be therapeutic interms of a partial or complete cure for a disease and/or adverse affectattributable to the disease. “Treatment,” as used herein, covers anytreatment of a disease in a mammal, particularly in a human, andincludes: (a) preventing the disease from occurring in a subject whichmay be predisposed to the disease but has not yet been diagnosed ashaving it; (b) inhibiting the disease, i.e., arresting its development;and (c) relieving the disease, i.e., causing regression of the disease.

The terms “individual,” “host,” “subject,” and “patient” are usedinterchangeably herein, and refer to a mammal, including, but notlimited to, murines, simians, humans, mammalian farm animals, mammaliansport animals, and mammalian pets.

As used herein, the term “a Type I interferon receptor agonist” refersto any naturally occurring or non-naturally occurring ligand of humanType I interferon receptor, which binds to and causes signaltransduction via the receptor. Type I interferon receptor agonistsinclude interferons, including naturally-occurring interferons, modifiedinterferons, synthetic interferons, pegylated interferons, fusionproteins comprising an interferon and a heterologous protein, shuffledinterferons; antibody specific for an interferon receptor; non-peptidechemical agonists; and the like.

As used herein, the term “Type II interferon receptor agonist” refers toany naturally occurring or non-naturally occurring ligand of human TypeII interferon receptor that binds to and causes signal transduction viathe receptor. Type II interferon receptor agonists include native humaninterferon-γ, recombinant IFN-γ species, glycosylated IFN-γ species,pegylated IFN-γ species, modified or variant IFN-γ species, IFN-γ fusionproteins, antibody agonists specific for the receptor, non-peptideagonists, and the like.

As used herein, the term “a Type III interferon receptor agonist” refersto any naturally occurring or non-naturally occurring ligand ofhumanIL-28 receptor α (“IL-28R”), the amino acid sequence of which isdescribed by Sheppard, et al., infra., that binds to and causes signaltransduction via the receptor.

As used herein, the term “interferon receptor agonist” refers to anyType I interferon receptor agonist, Type II interferon receptor agonist,or Type III interferon receptor agonist.

The term “dosing event” as used herein refers to administration of anantiviral agent to a patient in need thereof, which event may encompassone or more releases of an antiviral agent from a drug dispensingdevice. Thus, the term “dosing event,” as used herein, includes, but isnot limited to, installation of a continuous delivery device (e.g., apump or other controlled release injectable system); and a singlesubcutaneous injection followed by installation of a continuous deliverysystem.

“Continuous delivery” as used herein (e.g., in the context of“continuous delivery of a substance to a tissue”) is meant to refer tomovement of drug to a delivery site, e.g., into a tissue in a fashionthat provides for delivery of a desired amount of substance into thetissue over a selected period of time, where about the same quantity ofdrug is received by the patient each minute during the selected periodof time.

“Controlled release” as used herein (e.g., in the context of “controlleddrug release”) is meant to encompass release of substance (e.g., a TypeI or Type III interferon receptor agonist, e.g., IFN-α) at a selected orotherwise controllable rate, interval, and/or amount, which is notsubstantially influenced by the environment of use. “Controlled release”thus encompasses, but is not necessarily limited to, substantiallycontinuous delivery, and patterned delivery (e.g., intermittent deliveryover a period of time that is interrupted by regular or irregular timeintervals).

“Patterned” or “temporal” as used in the context of drug delivery ismeant delivery of drug in a pattern, generally a substantially regularpattern, over a pre-selected period of time (e.g., other than a periodassociated with, for example a bolus injection). “Patterned” or“temporal” drug delivery is meant to encompass delivery of drug at anincreasing, decreasing, substantially constant, or pulsatile, rate orrange of rates (e.g., amount of drug per unit time, or volume of drugformulation for a unit time), and further encompasses delivery that iscontinuous or substantially continuous, or chronic.

The term “controlled drug delivery device” is meant to encompass anydevice wherein the release (e.g., rate, timing of release) of a drug orother desired substance contained therein is controlled by or determinedby the device itself and not substantially influenced by the environmentof use, or releasing at a rate that is reproducible within theenvironment of use.

By “substantially continuous” as used in, for example, the context of“substantially continuous infusion” or “substantially continuousdelivery” is meant to refer to delivery of drug in a manner that issubstantially uninterrupted for a pre-selected period of drug delivery,where the quantity of drug received by the patient during any 8 hourinterval in the pre-selected period never falls to zero. Furthermore,“substantially continuous” drug delivery can also encompass delivery ofdrug at a substantially constant, pre-selected rate or range of rates(e.g., amount of drug per unit time, or volume of drug formulation for aunit time) that is substantially uninterrupted for a pre-selected periodof drug delivery.

By “substantially steady state” as used in the context of a biologicalparameter that may vary as a function of time, it is meant that thebiological parameter exhibits a substantially constant value over a timecourse, such that the area under the curve defined by the value of thebiological parameter as a function of time for any 8 hour period duringthe time course (AUC8 hr) is no more than about 20% above or about 20%below, and preferably no more than about 15% above or about 15% below,and more preferably no more than about 10% above or about 10% below, theaverage area under the curve of the biological parameter over an 8 hourperiod during the time course (AUC8 hr average). The AUC8 hr average isdefined as the quotient (q) of the area under the curve of thebiological parameter over the entirety of the time course (AUCtotal)divided by the number of 8 hour intervals in the time course (total/3days), i.e., q=(AUCtotal)/(total/3 days). For example, in the context ofa serum concentration of a drug, the serum concentration of the drug ismaintained at a substantially steady state during a time course when thearea under the curve of serum concentration of the drug over time forany 8 hour period during the time course (AUC8 hr) is no more than about20% above or about 20% below the average area under the curve of serumconcentration of the drug over an 8 hour period in the time course (AUC8hr average), i.e., the AUC8 hr is no more than 20% above or 20% belowthe AUC8 hr average for the serum concentration of the drug over thetime course.

The term “alkyl” used herein refers to a monovalent straight or branchedchain radical of from one to twenty carbon atoms, including, but notlimited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,tert-butyl, n-hexyl, and the like.

The term “halo” used herein refers to fluoro, chloro, bromo, or iodo.

The term “alkoxy” used herein refers to straight or branched chain alkylradical covalently bonded to the parent molecule through an —O— linkage.Examples of alkoxy groups include, but are not limited to, methoxy,ethoxy, propoxy, isopropoxy, butoxy, n-butoxy, sec-butoxy, t-butoxy andthe like.

The term “alkenyl” used herein refers to a monovalent straight orbranched chain radical of from two to twenty carbon atoms containing acarbon double bond including, but not limited to, 1-propenyl,2-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, and the like.

The term “alkynyl” used herein refers to a monovalent straight orbranched chain radical of from two to twenty carbon atoms containing acarbon triple bond including, but not limited to, 1-propynyl, 1-butynyl,2-butynyl, and the like.

The term “aryl” used herein refers to homocyclic aromatic radicalwhether fused or not fused. Examples of aryl groups include, but are notlimited to, phenyl, naphthyl, biphenyl, phenanthrenyl, naphthacenyl, andthe like.

The term “cycloalkyl” used herein refers to saturated aliphatic ringsystem radical having three to twenty carbon atoms including, but notlimited to, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, and thelike.

The term “cycloalkenyl” used herein refers to aliphatic ring systemradical having three to twenty carbon atoms having at least onecarbon-carbon double bond in the ring. Examples of cycloalkenyl groupsinclude, but are not limited to, cyclopropenyl, cyclopentenyl,cyclohexenyl, cycloheptenyl, and the like.

The term “polycycloalkyl” used herein refers to saturated aliphatic ringsystem radical having at least two rings that are fused with or withoutbridgehead carbons. Examples of polycycloalkyl groups include, but arenot limited to, bicyclo[4.4.0]decanyl, bicyclo[2.2.1]heptanyl,adamantyl, norbornyl, and the like.

The term “polycycloalkenyl” used herein refers to aliphatic ring systemradical having at least two rings that are fused with or withoutbridgehead carbons in which at least one of the rings has acarbon-carbon double bond. Examples of polycycloalkenyl groups include,but are not limited to, norbornylenyl, 1,1′-bicyclopentenyl, and thelike.

The term “polycyclic hydrocarbon” used herein refers to a ring systemradical in which all of the ring members are carbon atoms. Polycyclichydrocarbons can be aromatic or can contain less than the maximum numberof non-cumulative double bonds. Examples of polycyclic hydrocarboninclude, but are not limited to, naphthyl, dihydronaphthyl, indenyl,fluorenyl, and the like.

The term “heterocyclic” or “heterocyclyl” used herein refers to cyclicring system radical having at least one ring system in which one or morering atoms are not carbon, namely heteroatom. Heterocycles can benonaromatic or aromatic. Examples of heterocyclic groups include, butare not limited to, morpholinyl, tetrahydrofuranyl, dioxolanyl,pyrrolidinyl, oxazolyl, pyranyl, pyridyl, pyrimidinyl, pyrrolyl, and thelike.

The term “heteroaryl” used herein refers to heterocyclic group formallyderived from an arene by replacement of one or more methine and/orvinylene groups by trivalent or divalent heteroatoms, respectively, insuch a way as to maintain the aromatic system. Examples of heteroarylgroups include, but are not limited to, pyridyl, pyrrolyl, oxazolyl,indolyl, and the like.

The term “arylalkyl” used herein refers to one or more aryl groupsappended to an alkyl radical. Examples of arylalkyl groups include, butare not limited to, benzyl, phenethyl, phenpropyl, phenbutyl, and thelike.

The term “cycloalkylalkyl” used herein refers to one or more cycloalkylgroups appended to an alkyl radical. Examples of cycloalkylalkylinclude, but are not limited to, cyclohexylmethyl, cyclohexylethyl,cyclopentylmethyl, cyclopentylethyl, and the like.

The term “heteroarylalkyl” used herein refers to one or more heteroarylgroups appended to an alkyl radical. Examples of heteroarylalkylinclude, but are not limited to, pyridylmethyl, furanylmethyl,thiophenylethyl, and the like.

The term “heterocyclylalkyl” used herein refers to one or moreheterocyclyl groups appended to an alkyl radical. Examples ofheterocyclylalkyl include, but are not limited to, morpholinylmethyl,morpholinylethyl, morpholinylpropyl, tetrahydrofuranylmethyl,pyrrolidinylpropyl, and the like.

The term “aryloxy” used herein refers to an aryl radical covalentlybonded to the parent molecule through an —O— linkage.

The term “alkylthio” used herein refers to straight or branched chainalkyl radical covalently bonded to the parent molecule through an —S—linkage. Examples of alkoxy groups include, but are not limited to,methoxy, ethoxy, propoxy, isopropoxy, butoxy, n-butoxy, sec-butoxy,t-butoxy and the like.

The term “arylthio” used herein refers to an aryl radical covalentlybonded to the parent molecule through an —S— linkage.

The term “alkylamino” used herein refers to nitrogen radical with one ormore allyl groups attached thereto. Thus, monoalkylamino refers tonitrogen radical with one alkyl group attached thereto and dialkylaminorefers to nitrogen radical with two alkyl groups attached thereto.

The term “cyanoamino” used herein refers to nitrogen radical withnitrile group attached thereto.

The term “carbamyl” used herein refers to RNHCOO—.

The term “keto” and “carbonyl” used herein refers to C═O.

The term “carboxy” used herein refers to —COOH.

The term “sulfamyl” used herein refers to —SO₂NH₂.

The term “sulfonyl” used herein refers to —SO₂—.

The term “sulfinyl” used herein refers to —SO—.

The term “thiocarbonyl” used herein refers to C═S.

The term “thiocarboxy” used herein refers to CSOH.

As used herein, a radical indicates species with a single, unpairedelectron such that the species containing the radical can be covalentlybonded to another species. Hence, in this context, a radical is notnecessarily a free radical. Rather, a radical indicates a specificportion of a larger molecule. The term “radical” can be usedinterchangeably with the term “group.”

As used herein, a substituted group is derived from the unsubstitutedparent structure in which there has been an exchange of one or morehydrogen atoms for another atom or group. When substituted, thesubstituent group(s) is (are) one or more group(s) individually andindependently selected from C₁-C₆ alkyl, C₁-C₆ alkenyl, C₁-C₆ alkynyl,C₃-C₆ cycloalkyl, C₃-C₆ heterocycloalkyl (e.g., tetrahydrofuryl), aryl,heteroaryl, halo (e.g., chloro, bromo, iodo and fluoro), cyano, hydroxy,C₁-C₆ alkoxy, aryloxy, sulfhydryl (mercapto), C₁-C₆ alkylthio, arylthio,mono- and di-(C₁-C₆)alkyl amino, quaternary ammonium salts,amino(C₁-C₆)alkoxy, hydroxy(C₁-C₆)alkylamino, amino(C₁-C₆)alkylthio,cyanoamino, nitro, carbamyl, keto (oxy), carbonyl, carboxy, glycolyl,glycyl, hydrazino, guanyl, sulfamyl, sulfonyl, sulfinyl, thiocarbonyl,thiocarboxy, and combinations thereof. The protecting groups that canform the protective derivatives of the above substituents are known tothose of skill in the art and can be found in references such as Greeneand Wuts Protective Groups in Organic Synthesis; John Wiley and Sons:New York, 1999. Wherever a substituent is described as “optionallysubstituted” that substituent can be substituted with the abovesubstituents.

Asymmetric carbon atoms may be present in the compounds described. Allsuch isomers, including diastereomers and enantiomers, as well as themixtures thereof are intended to be included in the scope of the recitedcompound. In certain cases, compounds can exist in tautomeric forms. Alltautomeric forms are intended to be included in the scope of the recitedcompound. Likewise, when compounds contain an alkenyl or alkenylenegroup, there exists the possibility of cis- and trans-isomeric forms ofthe compounds. Both cis- and trans-isomers, as well as the mixtures ofcis- and trans-isomers, are contemplated. Thus, reference herein to acompound includes all of the aforementioned isomeric forms unless thecontext clearly dictates otherwise.

Various forms are included in the embodiments, including polymorphs,solvates, hydrates, conformers, salts, and prodrug derivatives. Apolymorph is a composition having the same chemical formula, but adifferent structure. A solvate is a composition formed by solvation (thecombination of solvent molecules with molecules or ions of the solute).A hydrate is a compound formed by an incorporation of water. A conformeris a structure that is a conformational isomer. Conformational isomerismis the phenomenon of molecules with the same structural formula butdifferent conformations (conformers) of atoms about a rotating bond.Salts of compounds can be prepared by methods known to those skilled inthe art. For example, salts of compounds can be prepared by reacting theappropriate base or acid with a stoichiometric equivalent of thecompound. A prodrug is a compound that undergoes biotransformation(chemical conversion) before exhibiting its pharmacological effects. Forexample, a prodrug can thus be viewed as a drug containing specializedprotective groups used in a transient manner to alter or to eliminateundesirable properties in the parent molecule. Thus, reference herein toa compound includes all of the aforementioned forms unless the contextclearly dictates otherwise.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range is encompassed within the embodiments. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges is also encompassed within the invention, subject to anyspecifically excluded limit in the stated range. Where the stated rangeincludes one or both of the limits, ranges excluding either both ofthose included limits are also included in the embodiments.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the embodiments belong. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the embodiments, the preferredmethods and materials are now described. All publications mentionedherein are incorporated herein by reference to disclose and describe themethods and/or materials in connection with which the publications arecited.

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “and,” and “the” include plural referents unless thecontext clearly dictates otherwise. Thus, for example, reference to “amethod” includes a plurality of such methods and reference to “a dose”includes reference to one or more doses and equivalents thereof known tothose skilled in the art, and so forth.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” is used herein to modify a numerical value above and below thestated value by a variance of 10%.

As used in this specification, whether in a transitional phrase or inthe body of the claim, the terms “comprise(s)” and “comprising” are tobe interpreted as having an open-ended meaning. That is, the terms areto be interpreted synonymously with the phrases “having at least” or“including at least”. When used in the context of a process, the term“comprising” means that the process includes at least the recited steps,but may include additional steps. When used in the context of acompound, composition or device, the term “comprising” means that thecompound, composition or device includes at least the recited featuresor components, but may also include additional features or components.

Reference is made herein in detail to specific embodiments of theinvention. While the invention will be described in conjunction withthese specific embodiments, it will be understood that it is notintended to limit the invention to such specific embodiments. On thecontrary, it is intended to cover alternatives, modifications, andequivalents as may be included within the spirit and scope of theinvention. In the description provided herein, numerous specific detailsare set forth in order to provide a thorough understanding of thepresent invention. The present invention may be practiced without someor all of these specific details. In other instances, well known processoperations have not been described in detail, in order not to obscurethe present invention.

The present embodiments provide Compounds 1-10 as described below, aswell as pharmaceutical compositions and formulations comprising anycompound of Compounds 1-10. The Compounds 1-10 may be referred to hereinin various ways, including “subject compound,” “NS3 inhibitor”, “NS3inhibitor compound”, “protease inhibitor”, “helicase inhibitor” andother similar terms that will be readily understood by those skilled inthe art. A subject compound is useful for modulating (e.g., inhibiting)the activity of NS3 protease and/or helicase, in vivo and/or ex vivo.For example, a subject compound is useful for treating HCV infection andother disorders, as discussed below.

Compositions

The present embodiments provide Compounds 1-10 having the chemicalstructures shown in Tables 1 and 2 below. Preferred embodiments providea method of treating a hepatitis C virus infection in an individual, themethod comprising administering to the individual an effective amount ofa composition comprising a preferred compound, e.g., a compositioncomprising at least one of Compounds 1-10.

Preferred embodiments provide a method of treating liver fibrosis in anindividual, the method comprising administering to the individual aneffective amount of a composition comprising a preferred compound.

Preferred embodiments provide a method of increasing liver function inan individual having a hepatitis C virus infection, the methodcomprising administering to the individual an effective amount of acomposition comprising a preferred compound.

The present embodiments further provide compositions, includingpharmaceutical compositions, comprising Compounds 1-10, and salts,esters, or other derivatives thereof. A subject pharmaceuticalcomposition comprises a subject compound; and a pharmaceuticallyacceptable excipient. A wide variety of pharmaceutically acceptableexcipients is known in the art and need not be discussed in detailherein. Pharmaceutically acceptable excipients have been amply describedin a variety of publications, including, for example, A. Gennaro (2000)“Remington: The Science and Practice of Pharmacy,” 20th edition,Lippincott, Williams, & Wilkins; Pharmaceutical Dosage Forms and DrugDelivery Systems (1999) H. C. Ansel et al., eds., 7^(th) ed.,Lippincott, Williams, & Wilkins; and Handbook of PharmaceuticalExcipients (2000) A. H. Kibbe et al., eds., 3^(rd) ed. Amer.Pharmaceutical Assoc.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,carriers or diluents, are readily available to the public. Moreover,pharmaceutically acceptable auxiliary substances, such as pH adjustingand buffering agents, tonicity adjusting agents, stabilizers, wettingagents and the like, are readily available to the public.

In many embodiments, a subject compound inhibits the enzymatic activityof a hepatitis virus C(HCV) NS3 protease. Whether a subject compoundinhibits HCV NS3 protease can be readily determined using any knownmethod. Typical methods involve a determination of whether an HCVpolyprotein or other polypeptide comprising an NS3 recognition site iscleaved by NS3 in the presence of the agent. In many embodiments, asubject compound inhibits NS3 enzymatic activity by at least about 10%,at least about 15%, at least about 20%, at least about 25%, at leastabout 30%, at least about 40%, at least about 50%, at least about 60%,at least about 70%, at least about 80%, or at least about 90%, or more,compared to the enzymatic activity of NS3 in the absence of thecompound.

In many embodiments, a subject compound inhibits enzymatic activity ofan HCV NS3 protease with an IC₅₀ of less than about 50 μM, e.g., asubject compound inhibits an HCV NS3 protease with an IC₅₀ of less thanabout 40 μM, less than about 25 μM, less than about 10 μM, less thanabout 1 μM, less than about 100 nM, less than about 80 nM, less thanabout 60 nM, less than about 50 nM, less than about 25 nM, less thanabout 10 nM, or less than about 1 nM, or less.

In many embodiments, a subject compound inhibits the enzymatic activityof a hepatitis virus C(HCV) NS3 helicase. Whether a subject compoundinhibits HCV NS3 helicase can be readily determined using any knownmethod. In many embodiments, a subject compound inhibits NS3 enzymaticactivity by at least about 10%, at least about 15%, at least about 20%,at least about 25%, at least about 30%, at least about 40%, at leastabout 50%, at least about 60%, at least about 70%, at least about 80%,or at least about 90%, or more, compared to the enzymatic activity ofNS3 in the absence of the compound.

In many embodiments, a subject compound inhibits HCV viral replication.For example, a subject compound inhibits HCV viral replication by atleast about 10%, at least about 15%, at least about 20%, at least about25%, at least about 30%, at least about 40%, at least about 50%, atleast about 60%, at least about 70%, at least about 80%, or at leastabout 90%, or more, compared to HCV viral replication in the absence ofthe compound. Whether a subject compound inhibits HCV viral replicationcan be determined using methods known in the art, including an in vitroviral replication assay.

In many embodiments, a subject compound inhibits the enzymatic activityof both HCV NS3 protease and HCV NS3 helicase. Whether a subjectcompound inhibits both HCV NS3 protease and HCV NS3 helicase may bereadily determined using any known method. In many embodiments, asubject compound inhibits both HCV NS3 protease and HCV NS3 helicaseenzymatic activity, each independently, by at least about 10%, at leastabout 15%, at least about 20%, at least about 25%, at least about 30%,at least about 40%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, or at least about 90%, or more, comparedto the HCV NS3 protease and HCV NS3 helicase enzymatic activity in theabsence of the compound.

Treating a Hepatitis Virus Infection

The methods and compositions described herein are generally useful intreatment of an of HCV infection.

Whether a subject method is effective in treating an HCV infection canbe determined by a reduction in viral load, a reduction in time toseroconversion (virus undetectable in patient serum), an increase in therate of sustained viral response to therapy, a reduction of morbidity ormortality in clinical outcomes, or other indicator of disease response.

In general, an effective amount of a compound described herein, e.g.,any one or more of Compounds 1-10, and optionally one or more additionalantiviral agents, is an amount that is effective to reduce viral load orachieve a sustained viral response to therapy.

Whether a subject method is effective in treating an HCV infection canbe determined by measuring viral load, or by measuring a parameterassociated with HCV infection, including, but not limited to, liverfibrosis, elevations in serum transaminase levels, and necroinflammatoryactivity in the liver. Indicators of liver fibrosis are discussed indetail below.

The method involves administering an effective amount of a compounddescribed herein, e.g., any one or more of Compounds 1-10, optionally incombination with an effective amount of one or more additional antiviralagents. In some embodiments, an effective amount of a compound describedherein, and optionally one or more additional antiviral agents, is anamount that is effective to reduce viral titers to undetectable levels,e.g., to about 1000 to about 5000, to about 500 to about 1000, or toabout 100 to about 500 genome copies/mL serum. In some embodiments, aneffective amount of a compound described herein, and optionally one ormore additional antiviral agents, is an amount that is effective toreduce viral load to lower than 100 genome copies/mL serum.

In some embodiments, an effective amount of a compound described herein,and optionally one or more additional antiviral agents, is an amountthat is effective to achieve a 1.5-log, a 2-log, a 2.5-log, a 3-log, a3.5-log, a 4-log, a 4.5-log, or a 5-log reduction in viral titer in theserum of the individual.

In many embodiments, an effective amount of a compound described herein,and optionally one or more additional antiviral agents, is an amountthat is effective to achieve a sustained viral response, e.g.,non-detectable or substantially non-detectable HCV RNA (e.g., less thanabout 500, less than about 400, less than about 200, or less than about100 genome copies per milliliter serum) is found in the patient's serumfor a period of at least about one month, at least about two months, atleast about three months, at least about four months, at least aboutfive months, or at least about six months following cessation oftherapy.

As noted above, whether a subject method is effective in treating an HCVinfection can be determined by measuring a parameter associated with HCVinfection, such as liver fibrosis. Methods of determining the extent ofliver fibrosis are discussed in detail below. In some embodiments, thelevel of a serum marker of liver fibrosis indicates the degree of liverfibrosis.

As one non-limiting example, levels of serum alanine aminotransferase(ALT) are measured, using standard assays. In general, an ALT level ofless than about 45 international units is considered normal. In someembodiments, an effective amount of a compound of formula I, andoptionally one or more additional antiviral agents, is an amounteffective to reduce ALT levels to less than about 45 IU/ml serum.

A therapeutically effective amount of a compound described herein, andoptionally one or more additional antiviral agents, is an amount that iseffective to reduce a serum level of a marker of liver fibrosis by atleast about 10%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, or at least about 80%, ormore, compared to the level of the marker in an untreated individual, orto a placebo-treated individual. Methods of measuring serum markersinclude immunological-based methods, e.g., enzyme-linked immunosorbentassays (ELISA), radioimmunoassays, and the like, using antibody specificfor a given serum marker.

In many embodiments, an effective amount of a compound described hereinand an additional antiviral agent is a synergistic amount. As usedherein, a “synergistic combination” or a “synergistic amount” of acompound described herein and an additional antiviral agent is acombined dosage that is more effective in the therapeutic orprophylactic treatment of an HCV infection than the incrementalimprovement in treatment outcome that could be predicted or expectedfrom a merely additive combination of (i) the therapeutic orprophylactic benefit of a compound described herein when administered atthat same dosage as a monotherapy and (ii) the therapeutic orprophylactic benefit of the additional antiviral agent when administeredat the same dosage as a monotherapy.

In some embodiments, a selected amount of a compound described hereinand a selected amount of an additional antiviral agent are effectivewhen used in combination therapy for a disease, but the selected amountof the compound described herein and/or the selected amount of theadditional antiviral agent is ineffective when used in monotherapy forthe disease. Thus, the embodiments encompass (1) regimens in which aselected amount of the additional antiviral agent enhances thetherapeutic benefit of a selected amount of the compound describedherein when used in combination therapy for a disease, where theselected amount of the additional antiviral agent provides notherapeutic benefit when used in monotherapy for the disease (2)regimens in which a selected amount of the compound described hereinenhances the therapeutic benefit of a selected amount of the additionalantiviral agent when used in combination therapy for a disease, wherethe selected amount of the compound described herein provides notherapeutic benefit when used in monotherapy for the disease and (3)regimens in which a selected amount of the compound described herein anda selected amount of the additional antiviral agent provide atherapeutic benefit when used in combination therapy for a disease,where each of the selected amounts of the compound described herein andthe additional antiviral agent, respectively, provides no therapeuticbenefit when used in monotherapy for the disease. As used herein, a“synergistically effective amount” of a compound described herein and anadditional antiviral agent, and its grammatical equivalents, shall beunderstood to include any regimen encompassed by any of (1)-(3) above.

Fibrosis

The embodiments provides methods for treating liver fibrosis (includingforms of liver fibrosis resulting from, or associated with, HCVinfection), generally involving administering a therapeutic amount of acompound described herein, and optionally one or more additionalantiviral agents. Effective amounts of any of Compounds 1-10, with andwithout one or more additional antiviral agents, as well as dosingregimens, are as discussed below.

Whether treatment with a compound described herein, and optionally oneor more additional antiviral agents, is effective in reducing liverfibrosis is determined by any of a number of well-established techniquesfor measuring liver fibrosis and liver function. Liver fibrosisreduction is determined by analyzing a liver biopsy sample. An analysisof a liver biopsy comprises assessments of two major components:necroinflammation assessed by “grade” as a measure of the severity andongoing disease activity, and the lesions of fibrosis and parenchymal orvascular remodeling as assessed by “stage” as being reflective oflong-term disease progression. See, e.g., Brunt (2000) Hepatol.31:241-246; and METAVIR (1994) Hepatology 20:15-20. Based on analysis ofthe liver biopsy, a score is assigned. A number of standardized scoringsystems exist which provide a quantitative assessment of the degree andseverity of fibrosis. These include the METAVIR, Knodell, Scheuer,Ludwig, and Ishak scoring systems.

The METAVIR scoring system is based on an analysis of various featuresof a liver biopsy, including fibrosis (portal fibrosis, centrilobularfibrosis, and cirrhosis); necrosis (piecemeal and lobular necrosis,acidophilic retraction, and ballooning degeneration); inflammation(portal tract inflammation, portal lymphoid aggregates, and distributionof portal inflammation); bile duct changes; and the Knodell index(scores of periportal necrosis, lobular necrosis, portal inflammation,fibrosis, and overall disease activity). The definitions of each stagein the METAVIR system are as follows: score: 0, no fibrosis; score: 1,stellate enlargement of portal tract but without septa formation; score:2, enlargement of portal tract with rare septa formation; score: 3,numerous septa without cirrhosis; and score: 4, cirrhosis.

Knodell's scoring system, also called the Hepatitis Activity Index,classifies specimens based on scores in four categories of histologicfeatures: I. Periportal and/or bridging necrosis; II. Intralobulardegeneration and focal necrosis; III. Portal inflammation; and IV.Fibrosis. In the Knodell staging system, scores are as follows: score:0, no fibrosis; score: 1, mild fibrosis (fibrous portal expansion);score: 2, moderate fibrosis; score: 3, severe fibrosis (bridgingfibrosis); and score: 4, cirrhosis. The higher the score, the moresevere the liver tissue damage. Knodell (1981) Hepatol. 1:431.

In the Scheuer scoring system scores are as follows: score: 0, nofibrosis; score: 1, enlarged, fibrotic portal tracts; score: 2,periportal or portal-portal septa, but intact architecture; score: 3,fibrosis with architectural distortion, but no obvious cirrhosis; score:4, probable or definite cirrhosis. Scheuer (1991) J. Hepatol. 13:372.

The Ishak scoring system is described in Ishak (1995) J. Hepatol.22:696-699. Stage 0, No fibrosis; Stage 1, Fibrous expansion of someportal areas, with or without short fibrous septa; stage 2, Fibrousexpansion of most portal areas, with or without short fibrous septa;stage 3, Fibrous expansion of most portal areas with occasional portalto portal (P-P) bridging; stage 4, Fibrous expansion of portal areaswith marked bridging (P-P) as well as portal-central (P-C); stage 5,Marked bridging (P-P and/or P-C) with occasional nodules (incompletecirrhosis); stage 6, Cirrhosis, probable or definite.

The benefit of anti-fibrotic therapy can also be measured and assessedby using the Child-Pugh scoring system which comprises a multicomponentpoint system based upon abnormalities in serum bilirubin level, serumalbumin level, prothrombin time, the presence and severity of ascites,and the presence and severity of encephalopathy. Based upon the presenceand severity of abnormality of these parameters, patients may be placedin one of three categories of increasing severity of clinical disease:A, B, or C.

In some embodiments, a therapeutically effective amount of a compound offormula I, and optionally one or more additional antiviral agents, is anamount that effects a change of one unit or more in the fibrosis stagebased on pre- and post-therapy liver biopsies. In particularembodiments, a therapeutically effective amount of a compound of formulaI, and optionally one or more additional antiviral agents, reduces liverfibrosis by at least one unit in the METAVIR, the Knodell, the Scheuer,the Ludwig, or the Ishak scoring system.

Secondary, or indirect, indices of liver function can also be used toevaluate the efficacy of treatment with a compound described herein.Morphometric computerized semi-automated assessment of the quantitativedegree of liver fibrosis based upon specific staining of collagen and/orserum markers of liver fibrosis can also be measured as an indication ofthe efficacy of a subject treatment method. Secondary indices of liverfunction include, but are not limited to, serum transaminase levels,prothrombin time, bilirubin, platelet count, portal pressure, albuminlevel, and assessment of the Child-Pugh score.

An effective amount of a compound described herein, and optionally oneor more additional antiviral agents, is an amount that is effective toincrease an index of liver function by at least about 10%, at leastabout 20%, at least about 25%, at least about 30%, at least about 35%,at least about 40%, at least about 45%, at least about 50%, at leastabout 55%, at least about 60%, at least about 65%, at least about 70%,at least about 75%, or at least about 80%, or more, compared to theindex of liver function in an untreated individual, or to aplacebo-treated individual. Those skilled in the art can readily measuresuch indices of liver function, using standard assay methods, many ofwhich are commercially available, and are used routinely in clinicalsettings.

Serum markers of liver fibrosis can also be measured as an indication ofthe efficacy of a subject treatment method. Serum markers of liverfibrosis include, but are not limited to, hyaluronate, N-terminalprocollagen III peptide, 7S domain of type IV collagen, C-terminalprocollagen I peptide, and laminin. Additional biochemical markers ofliver fibrosis include α-2-macroglobulin, haptoglobin, gamma globulin,apolipoprotein A, and gamma glutamyl transpeptidase.

A therapeutically effective amount of a compound described herein, andoptionally one or more additional antiviral agents, is an amount that iseffective to reduce a serum level of a marker of liver fibrosis by atleast about 10%, at least about 20%, at least about 25%, at least about30%, at least about 35%, at least about 40%, at least about 45%, atleast about 50%, at least about 55%, at least about 60%, at least about65%, at least about 70%, at least about 75%, or at least about 80%, ormore, compared to the level of the marker in an untreated individual, orto a placebo-treated individual. Those skilled in the art can readilymeasure such serum markers of liver fibrosis, using standard assaymethods, many of which are commercially available, and are usedroutinely in clinical settings. Methods of measuring serum markersinclude immunological-based methods, e.g., enzyme-linked immunosorbentassays (ELISA), radioimmunoassays, and the like, using antibody specificfor a given serum marker.

Quantitative tests of functional liver reserve can also be used toassess the efficacy of treatment with an interferon receptor agonist andpirfenidone (or a pirfenidone analog). These include: indocyanine greenclearance (ICG), galactose elimination capacity (GEC), aminopyrinebreath test (ABT), antipyrine clearance, monoethylglycine-xylidide(MEG-X) clearance, and caffeine clearance.

As used herein, a “complication associated with cirrhosis of the liver”refers to a disorder that is a sequellae of decompensated liver disease,i.e., or occurs subsequently to and as a result of development of liverfibrosis, and includes, but it not limited to, development of ascites,variceal bleeding, portal hypertension, jaundice, progressive liverinsufficiency, encephalopathy, hepatocellular carcinoma, liver failurerequiring liver transplantation, and liver-related mortality.

A therapeutically effective amount of a compound described herein, andoptionally one or more additional antiviral agents, is an amount that iseffective in reducing the incidence (e.g., the likelihood that anindividual will develop) of a disorder associated with cirrhosis of theliver by at least about 10%, at least about 20%, at least about 25%, atleast about 30%, at least about 35%, at least about 40%, at least about45%, at least about 50%, at least about 55%, at least about 60%, atleast about 65%, at least about 70%, at least about 75%, or at leastabout 80%, or more, compared to an untreated individual, or to aplacebo-treated individual.

Whether treatment with a compound described herein, and optionally oneor more additional antiviral agents, is effective in reducing theincidence of a disorder associated with cirrhosis of the liver canreadily be determined by those skilled in the art.

Reduction in liver fibrosis increases liver function. Thus, theembodiments provide methods for increasing liver function, generallyinvolving administering a therapeutically effective amount of a compounddescribed herein, and optionally one or more additional antiviralagents. Liver functions include, but are not limited to, synthesis ofproteins such as serum proteins (e.g., albumin, clotting factors,alkaline phosphatase, aminotransferases (e.g., alanine transaminase,aspartate transaminase), 5′-nucleosidase, γ-glutaminyltranspeptidase,etc.), synthesis of bilirubin, synthesis of cholesterol, and synthesisof bile acids; a liver metabolic function, including, but not limitedto, carbohydrate metabolism, amino acid and ammonia metabolism, hormonemetabolism, and lipid metabolism; detoxification of exogenous drugs; ahemodynamic function, including splanchnic and portal hemodynamics; andthe like.

Whether a liver function is increased is readily ascertainable by thoseskilled in the art, using well-established tests of liver function.Thus, synthesis of markers of liver function such as albumin, alkalinephosphatase, alanine transaminase, aspartate transaminase, bilirubin,and the like, can be assessed by measuring the level of these markers inthe serum, using standard immunological and enzymatic assays. Splanchniccirculation and portal hemodynamics can be measured by portal wedgepressure and/or resistance using standard methods. Metabolic functionscan be measured by measuring the level of ammonia in the serum.

Whether serum proteins normally secreted by the liver are in the normalrange can be determined by measuring the levels of such proteins, usingstandard immunological and enzymatic assays. Those skilled in the artknow the normal ranges for such serum proteins. The following arenon-limiting examples. The normal level of alanine transaminase is about45 IU per milliliter of serum. The normal range of aspartatetransaminase is from about 5 to about 40 units per liter of serum.Bilirubin is measured using standard assays. Normal bilirubin levels areusually less than about 1.2 mg/dL. Serum albumin levels are measuredusing standard assays. Normal levels of serum albumin are in the rangeof from about 35 to about 55 g/L. Prolongation of prothrombin time ismeasured using standard assays. Normal prothrombin time is less thanabout 4 seconds longer than control.

A therapeutically effective amount of a compound described herein, andoptionally one or more additional antiviral agents, is one that iseffective to increase liver function by at least about 10%, at leastabout 20%, at least about 30%, at least about 40%, at least about 50%,at least about 60%, at least about 70%, at least about 80%, or more. Forexample, a therapeutically effective amount of a compound describedherein, and optionally one or more additional antiviral agents, is anamount effective to reduce an elevated level of a serum marker of liverfunction by at least about 10%, at least about 20%, at least about 30%,at least about 40%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, or more, or to reduce the level of theserum marker of liver function to within a normal range. Atherapeutically effective amount of a compound described herein, andoptionally one or more additional antiviral agents, is also an amounteffective to increase a reduced level of a serum marker of liverfunction by at least about 10%, at least about 20%, at least about 30%,at least about 40%, at least about 50%, at least about 60%, at leastabout 70%, at least about 80%, or more, or to increase the level of theserum marker of liver function to within a normal range.

Dosages, Formulations, and Routes of Administration

In the subject methods, the active agent(s) (e.g., a compound orcompounds described herein, and optionally one or more additionalantiviral agents) may be administered to the host using any convenientmeans capable of resulting in the desired therapeutic effect. Thus, theagent can be incorporated into a variety of formulations for therapeuticadministration. More particularly, the agents of the embodiments may beformulated into pharmaceutical compositions by combination withappropriate, pharmaceutically acceptable carriers or diluents, and maybe formulated into preparations in solid, semi-solid, liquid or gaseousforms, such as tablets, capsules, powders, granules, ointments,solutions, suppositories, injections, inhalants and aerosols.

Formulations

The above-discussed active agent(s) (e.g., the Compounds 1-10 set forthin Tables 1-2 below) may be formulated using well-known reagents andmethods. Compositions are provided in formulation with apharmaceutically acceptable excipient(s). A wide variety ofpharmaceutically acceptable excipients are known in the art and need notbe discussed in detail herein. Pharmaceutically acceptable excipientshave been amply described in a variety of publications, including, forexample, A. Gennaro (2000) “Remington: The Science and Practice ofPharmacy,” 20th edition, Lippincott, Williams, & Wilkins; PharmaceuticalDosage Forms and Drug Delivery Systems (1999) H. C. Ansel et al., eds.,7^(th) ed., Lippincott, Williams, & Wilkins; and Handbook ofPharmaceutical Excipients (2000) A. H. Kibbe et al., eds., 3^(rd) ed.Amer. Pharmaceutical Assoc.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,carriers or diluents, are readily available to the public. Moreover,pharmaceutically acceptable auxiliary substances, such as pH adjustingand buffering agents, tonicity adjusting agents, stabilizers, wettingagents and the like, are readily available to the public.

In some embodiments, an agent is formulated in an aqueous buffer.Suitable aqueous buffers include, but are not limited to, acetate,succinate, citrate, and phosphate buffers varying in strengths fromabout 5 mM to about 100 mM. In some embodiments, the aqueous bufferincludes reagents that provide for an isotonic solution. Such reagentsinclude, but are not limited to, sodium chloride; and sugars e.g.,mannitol, dextrose, sucrose, and the like. In some embodiments, theaqueous buffer further includes a non-ionic surfactant such aspolysorbate 20 or 80. Optionally the formulations may further include apreservative. Suitable preservatives include, but are not limited to, abenzyl alcohol, phenol, chlorobutanol, benzalkonium chloride, and thelike. In many cases, the formulation is stored at about 4° C.Formulations may also be lyophilized, in which case they generallyinclude cryoprotectants such as sucrose, trehalose, lactose, maltose,mannitol, and the like. Lyophilized formulations can be stored overextended periods of time, even at ambient temperatures.

As such, administration of the agents can be achieved in various ways,including oral, buccal, rectal, parenteral, intraperitoneal,intradermal, subcutaneous, intramuscular, transdermal, intratracheal,etc., administration. In many embodiments, administration is by bolusinjection, e.g., subcutaneous bolus injection, intramuscular bolusinjection, and the like.

The pharmaceutical compositions of the embodiments can be administeredorally, parenterally or via an implanted reservoir. Oral administrationor administration by injection is preferred.

Subcutaneous administration of a pharmaceutical composition of theembodiments is accomplished using standard methods and devices, e.g.,needle and syringe, a subcutaneous injection port delivery system, andthe like. See, e.g., U.S. Pat. Nos. 3,547,119; 4,755,173; 4,531,937;4,311,137; and 6,017,328. A combination of a subcutaneous injection portand a device for administration of a pharmaceutical composition of theembodiments to a patient through the port is referred to herein as “asubcutaneous injection port delivery system.” In many embodiments,subcutaneous administration is achieved by bolus delivery by needle andsyringe.

In pharmaceutical dosage forms, the agents may be administered in theform of their pharmaceutically acceptable salts, or they may also beused alone or in appropriate association, as well as in combination,with other pharmaceutically active compounds. The following methods andexcipients are merely exemplary and are in no way limiting.

For oral preparations, the agents can be used alone or in combinationwith appropriate additives to make tablets, powders, granules orcapsules, for example, with conventional additives, such as lactose,mannitol, corn starch or potato starch; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, corn starch orgelatins; with disintegrators, such as corn starch, potato starch orsodium carboxymethylcellulose; with lubricants, such as talc ormagnesium stearate; and if desired, with diluents, buffering agents,moistening agents, preservatives and flavoring agents.

The agents can be formulated into preparations for injection bydissolving, suspending or emulsifying them in an aqueous or nonaqueoussolvent, such as vegetable or other similar oils, synthetic aliphaticacid glycerides, esters of higher aliphatic acids or propylene glycol;and if desired, with conventional additives such as solubilizers,isotonic agents, suspending agents, emulsifying agents, stabilizers andpreservatives.

Furthermore, the agents can be made into suppositories by mixing with avariety of bases such as emulsifying bases or water-soluble bases. Thecompounds of the embodiments can be administered rectally via asuppository. The suppository can include vehicles such as cocoa butter,carbowaxes and polyethylene glycols, which melt at body temperature, yetare solidified at room temperature.

Unit dosage forms for oral or rectal administration such as syrups,elixirs, and suspensions may be provided wherein each dosage unit, forexample, teaspoonful, tablespoonful, tablet or suppository, contains apredetermined amount of the composition containing one or moreinhibitors. Similarly, unit dosage forms for injection or intravenousadministration may comprise the inhibitor(s) in a composition as asolution in sterile water, normal saline or another pharmaceuticallyacceptable carrier.

The term “unit dosage form,” as used herein, refers to physicallydiscrete units suitable as unitary dosages for human and animalsubjects, each unit containing a predetermined quantity of compounds ofthe embodiments calculated in an amount sufficient to produce thedesired effect in association with a pharmaceutically acceptablediluent, carrier or vehicle. The specifications for the novel unitdosage forms of the embodiments depend on the particular compoundemployed and the effect to be achieved, and the pharmacodynamicsassociated with each compound in the host.

The pharmaceutically acceptable excipients, such as vehicles, adjuvants,carriers or diluents, are readily available to the public. Moreover,pharmaceutically acceptable auxiliary substances, such as pH adjustingand buffering agents, tonicity adjusting agents, stabilizers, wettingagents and the like, are readily available to the public.

Other Antiviral or Antifibrotic Agents

As discussed above, a subject method will in some embodiments be carriedout by administering an NS3 inhibitor that is a compound describedherein, and optionally one or more additional antiviral agent(s).

In some embodiments, the method further includes administration of oneor more interferon receptor agonist(s). Interferon receptor agonists aredescribed above.

In other embodiments, the method further includes administration ofpirfenidone or a pirfenidone analog. Pirfenidone and pirfenidone analogsare described above.

Additional antiviral agents that are suitable for use in combinationtherapy include, but are not limited to, nucleotide and nucleosideanalogs. Non-limiting examples include azidothymidine (AZT)(zidovudine), and analogs and derivatives thereof; 2′,3′-dideoxyinosine(DDI) (didanosine), and analogs and derivatives thereof;2′,3′-dideoxycytidine (DDC) (dideoxycytidine), and analogs andderivatives thereof; 2′3,′-didehydro-2′,3′-dideoxythymidine (D4T)(stavudine), and analogs and derivatives thereof; combivir; abacavir;adefovir dipoxil; cidofovir; ribavirin; ribavirin analogs; and the like.

In some embodiments, the method further includes administration ofribavirin. Ribavirin,1-β-D-ribofuranosyl-1H-1,2,4-triazole-3-carboxamide, available from ICNPharmaceuticals, Inc., Costa Mesa, Calif., is described in the MerckIndex, compound No. 8199, Eleventh Edition. Its manufacture andformulation is described in U.S. Pat. No. 4,211,771. Some embodimentsalso involve use of derivatives of ribavirin (see, e.g., U.S. Pat. No.6,277,830). The ribavirin may be administered orally in capsule ortablet form, or in the same or different administration form and in thesame or different route as the interferon receptor agonist. Of course,other types of administration of both medicaments, as they becomeavailable are contemplated, such as by nasal spray, transdermally,intravenously, by suppository, by sustained release dosage form, etc.Any form of administration will work so long as the proper dosages aredelivered without destroying the active ingredient.

In some embodiments, an additional antiviral agent is administeredduring the entire course of NS3 inhibitor compound treatment. In otherembodiments, an additional antiviral agent is administered for a periodof time that is overlapping with that of the NS3 inhibitor compoundtreatment, e.g., the additional antiviral agent treatment can beginbefore the NS3 inhibitor compound treatment begins and end before theNS3 inhibitor compound treatment ends; the additional antiviral agenttreatment can begin after the NS3 inhibitor compound treatment beginsand end after the NS3 inhibitor compound treatment ends; the additionalantiviral agent treatment can begin after the NS3 inhibitor compoundtreatment begins and end before the NS3 inhibitor compound treatmentends; or the additional antiviral agent treatment can begin before theNS3 inhibitor compound treatment begins and end after the NS3 inhibitorcompound treatment ends.

Methods of Treatment

Monotherapies

The NS3 inhibitor compounds described herein may be used in acute orchronic therapy for HCV disease. In many embodiments, the NS3 inhibitorcompound is administered for a period of about 1 day to about 7 days, orabout 1 week to about 2 weeks, or about 2 weeks to about 3 weeks, orabout 3 weeks to about 4 weeks, or about 1 month to about 2 months, orabout 3 months to about 4 months, or about 4 months to about 6 months,or about 6 months to about 8 months, or about 8 months to about 12months, or at least one year, and may be administered over longerperiods of time. The NS3 inhibitor compound can be administered 5 timesper day, 4 times per day, tid, bid, qd, qod, biw, tiw, qw, qow, threetimes per month, or once monthly. In other embodiments, the NS3inhibitor compound is administered as a continuous infusion.

In many embodiments, an NS3 inhibitor compound of the embodiments isadministered orally.

In connection with the above-described methods for the treatment of HCVdisease in a patient, an NS3 inhibitor compound as described herein maybe administered to the patient at a dosage from about 0.01 mg to about100 mg/kg patient bodyweight per day, in 1 to 5 divided doses per day.In some embodiments, the NS3 inhibitor compound is administered at adosage of about 0.5 mg to about 75 mg/kg patient bodyweight per day, in1 to 5 divided doses per day.

The amount of active ingredient that may be combined with carriermaterials to produce a dosage form can vary depending on the host to betreated and the particular mode of administration. A typicalpharmaceutical preparation can contain from about 5% to about 95% activeingredient (w/w). In other embodiments, the pharmaceutical preparationcan contain from about 20% to about 80% active ingredient.

Those of skill will readily appreciate that dose levels can vary as afunction of the specific NS3 inhibitor compound, the severity of thesymptoms and the susceptibility of the subject to side effects.Preferred dosages for a given NS3 inhibitor compound are readilydeterminable by those of skill in the art by a variety of means. Apreferred means is to measure the physiological potency of a giveninterferon receptor agonist.

In many embodiments, multiple doses of NS3 inhibitor compound areadministered. For example, an NS3 inhibitor compound is administeredonce per month, twice per month, three times per month, every other week(qow), once per week (qw), twice per week (biw), three times per week(tiw), four times per week, five times per week, six times per week,every other day (qod), daily (qd), twice a day (qid), or three times aday (tid), over a period of time ranging from about one day to about oneweek, from about two weeks to about four weeks, from about one month toabout two months, from about two months to about four months, from aboutfour months to about six months, from about six months to about eightmonths, from about eight months to about 1 year, from about 1 year toabout 2 years, or from about 2 years to about 4 years, or more.

Combination Therapies with Ribavirin

In some embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of ribavirin. Ribavirin can be administered indosages of about 400 mg, about 800 mg, about 1000 mg, or about 1200 mgper day.

One embodiment provides any of the above-described methods modified toinclude co-administering to the patient a therapeutically effectiveamount of ribavirin for the duration of the desired course of NS3inhibitor compound treatment.

Another embodiment provides any of the above-described methods modifiedto include co-administering to the patient about 800 mg to about 1200 mgribavirin orally per day for the duration of the desired course of NS3inhibitor compound treatment. In another embodiment, any of theabove-described methods may be modified to include co-administering tothe patient (a) 1000 mg ribavirin orally per day if the patient has abody weight less than 75 kg or (b) 1200 mg ribavirin orally per day ifthe patient has a body weight greater than or equal to 75 kg, where thedaily dosage of ribavirin is optionally divided into to 2 doses for theduration of the desired course of NS3 inhibitor compound treatment.

Combination Therapies with Levovirin

In some embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of levovirin. Levovirin is generallyadministered in an amount ranging from about 30 mg to about 60 mg, fromabout 60 mg to about 125 mg, from about 125 mg to about 200 mg, fromabout 200 mg to about 300 gm, from about 300 mg to about 400 mg, fromabout 400 mg to about 1200 mg, from about 600 mg to about 1000 mg, orfrom about 700 to about 900 mg per day, or about 10 mg/kg body weightper day. In some embodiments, levovirin is administered orally indosages of about 400, about 800, about 1000, or about 1200 mg per dayfor the desired course of NS3 inhibitor compound treatment.

Combination Therapies with Viramidine

In some embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of viramidine. Viramidine is generallyadministered in an amount ranging from about 30 mg to about 60 mg, fromabout 60 mg to about 125 mg, from about 125 mg to about 200 mg, fromabout 200 mg to about 300 gm, from about 300 mg to about 400 mg, fromabout 400 mg to about 1200 mg, from about 600 mg to about 1000 mg, orfrom about 700 to about 900 mg per day, or about 10 mg/kg body weightper day. In some embodiments, viramidine is administered orally indosages of about 800, or about 1600 mg per day for the desired course ofNS3 inhibitor compound treatment.

Combination Therapies with Alpha-Glucosidase Inhibitors

Suitable α-glucosidase inhibitors include any of the above-describedimino-sugars, including long-alkyl chain derivatives of imino sugars asdisclosed in U.S. Patent Publication No. 2004/0110795; inhibitors ofendoplasmic reticulum-associated α-glucosidases; inhibitors of membranebound α-glucosidase; miglitol (Glyset®), and active derivatives, andanalogs thereof; and acarbose (Precose®), and active derivatives, andanalogs thereof.

In many embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of an α-glucosidase inhibitor administered for aperiod of about 1 day to about 7 days, or about 1 week to about 2 weeks,or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks, orabout 1 month to about 2 months, or about 3 months to about 4 months, orabout 4 months to about 6 months, or about 6 months to about 8 months,or about 8 months to about 12 months, or at least one year, and may beadministered over longer periods of time.

An α-glucosidase inhibitor can be administered 5 times per day, 4 timesper day, tid (three times daily), bid, qd, qod, biw, tiw, qw, qow, threetimes per month, or once monthly. In other embodiments, an α-glucosidaseinhibitor is administered as a continuous infusion.

In many embodiments, an α-glucosidase inhibitor is administered orally.

In connection with the above-described methods for the treatment of aflavivirus infection, treatment of HCV infection, and treatment of liverfibrosis that occurs as a result of an HCV infection, the methodsprovide for combination therapy comprising administering an NS3inhibitor compound as described above, and an effective amount ofα-glucosidase inhibitor administered to the patient at a dosage of fromabout 10 mg per day to about 600 mg per day in divided doses, e.g., fromabout 10 mg per day to about 30 mg per day, from about 30 mg per day toabout 60 mg per day, from about 60 mg per day to about 75 mg per day,from about 75 mg per day to about 90 mg per day, from about 90 mg perday to about 120 mg per day, from about 120 mg per day to about 150 mgper day, from about 150 mg per day to about 180 mg per day, from about180 mg per day to about 210 mg per day, from about 210 mg per day toabout 240 mg per day, from about 240 mg per day to about 270 mg per day,from about 270 mg per day to about 300 mg per day, from about 300 mg perday to about 360 mg per day, from about 360 mg per day to about 420 mgper day, from about 420 mg per day to about 480 mg per day, or fromabout 480 mg to about 600 mg per day.

In some embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of α-glucosidase inhibitor administered in adosage of about 10 mg three times daily. In some embodiments, anα-glucosidase inhibitor is administered in a dosage of about 15 mg threetimes daily. In some embodiments, an α-glucosidase inhibitor isadministered in a dosage of about 20 mg three times daily. In someembodiments, an α-glucosidase inhibitor is administered in a dosage ofabout 25 mg three times daily. In some embodiments, an α-glucosidaseinhibitor is administered in a dosage of about 30 mg three times daily.In some embodiments, an α-glucosidase inhibitor is administered in adosage of about 40 mg three times daily. In some embodiments, anα-glucosidase inhibitor is administered in a dosage of about 50 mg threetimes daily. In some embodiments, an α-glucosidase inhibitor isadministered in a dosage of about 100 mg three times daily. In someembodiments, an α-glucosidase inhibitor is administered in a dosage ofabout 75 mg per day to about 150 mg per day in two or three divideddoses, where the individual weighs 60 kg or less. In some embodiments,an α-glucosidase inhibitor is administered in a dosage of about 75 mgper day to about 300 mg per day in two or three divided doses, where theindividual weighs 60 kg or more.

The amount of active ingredient (e.g., α-glucosidase inhibitor) that maybe combined with carrier materials to produce a dosage form can varydepending on the host to be treated and the particular mode ofadministration. A typical pharmaceutical preparation can contain fromabout 5% to about 95% active ingredient (w/w). In other embodiments, thepharmaceutical preparation can contain from about 20% to about 80%active ingredient.

Those of skill will readily appreciate that dose levels can vary as afunction of the specific α-glucosidase inhibitor, the severity of thesymptoms and the susceptibility of the subject to side effects.Preferred dosages for a given α-glucosidase inhibitor are readilydeterminable by those of skill in the art by a variety of means. Atypical means is to measure the physiological potency of a given activeagent.

In many embodiments, multiple doses of an α-glucosidase inhibitor areadministered. For example, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of α-glucosidase inhibitor administered once permonth, twice per month, three times per month, every other week (qow),once per week (qw), twice per week (biw), three times per week (tiw),four times per week, five times per week, six times per week, everyother day (qod), daily (qd), twice a day (qid), or three times a day(tid), over a period of time ranging from about one day to about oneweek, from about two weeks to about four weeks, from about one month toabout two months, from about two months to about four months, from aboutfour months to about six months, from about six months to about eightmonths, from about eight months to about 1 year, from about 1 year toabout 2 years, or from about 2 years to about 4 years, or more.

Combination Therapies with Thymosin-α

In some embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of thymosin-α. Thymosin-α (Zadaxin™) isgenerally administered by subcutaneous injection. Thymosin-α can beadministered tid, bid, qd, qod, biw, tiw, qw, qow, three times permonth, once monthly, substantially continuously, or continuously for thedesired course of NS3 inhibitor compound treatment. In many embodiments,thymosin-α is administered twice per week for the desired course of NS3inhibitor compound treatment. Effective dosages of thymosin-α range fromabout 0.5 mg to about 5 mg, e.g., from about 0.5 mg to about 1.0 mg,from about 1.0 mg to about 1.5 mg, from about 1.5 mg to about 2.0 mg,from about 2.0 mg to about 2.5 mg, from about 2.5 mg to about 3.0 mg,from about 3.0 mg to about 3.5 mg, from about 3.5 mg to about 4.0 mg,from about 4.0 mg to about 4.5 mg, or from about 4.5 mg to about 5.0 mg.In particular embodiments, thymosin-α is administered in dosagescontaining an amount of 1.0 mg or 1.6 mg.

Thymosin-α can be administered over a period of time ranging from aboutone day to about one week, from about two weeks to about four weeks,from about one month to about two months, from about two months to aboutfour months, from about four months to about six months, from about sixmonths to about eight months, from about eight months to about 1 year,from about 1 year to about 2 years, or from about 2 years to about 4years, or more. In one embodiment, thymosin-α is administered for thedesired course of NS3 inhibitor compound treatment.

Combination Therapies with Interferon(s)

In many embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of an interferon receptor agonist. In someembodiments, a compound described herein and a Type I or III interferonreceptor agonist are co-administered in the treatment methods describedherein. Type I interferon receptor agonists suitable for use hereininclude any interferon-α (IFN-α). In certain embodiments, theinterferon-α is a PEGylated interferon-α. In certain other embodiments,the interferon-α is a consensus interferon, such as INFERGEN® interferonalfacon-1. In still other embodiments, the interferon-α is a monoPEG (30kD, linear)-ylated consensus interferon.

Effective dosages of an IFN-α range from about 3 μg to about 27 μg, fromabout 3 MU to about 10 MU, from about 90 μg to about 180 μg, or fromabout 18 μg to about 90 μg. Effective dosages of Infergen® consensusIFN-α include about 3 μg, about 6 μg, about 9 μg, about 12 μg, about 15μg, about 18 μg, about 21 μg, about 24 μg, about 27 μg, or about 30 μg,of drug per dose. Effective dosages of IFN-α2a and IFN-α2b range from 3million Units (MU) to 10 MU per dose. Effective dosages ofPEGASYS®PEGylated IFN-α2a contain an amount of about 90 fig to 270 μg,or about 180 μg, of drug per dose. Effective dosages ofPEG-INTRON®PEGylated IFN-α2b contain an amount of about 0.5 μg to 3.0 μgof drug per kg of body weight per dose. Effective dosages of PEGylatedconsensus interferon (PEG-CIFN) contain an amount of about 18 μg toabout 90 μg, or from about 27 μg to about 60 μg, or about 45 μg, of CIFNamino acid weight per dose of PEG-CIFN. Effective dosages of monoPEG (30kD, linear)-ylated CIFN contain an amount of about 45 μg to about 270μg, or about 60 μg to about 180 μg, or about 90 μg to about 120 μg, ofdrug per dose. IFN-α can be administered daily, every other day, once aweek, three times a week, every other week, three times per month, oncemonthly, substantially continuously or continuously.

In many embodiments, the Type I or Type III interferon receptor agonistand/or the Type II interferon receptor agonist is administered for aperiod of about 1 day to about 7 days, or about 1 week to about 2 weeks,or about 2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks, orabout 1 month to about 2 months, or about 3 months to about 4 months, orabout 4 months to about 6 months, or about 6 months to about 8 months,or about 8 months to about 12 months, or at least one year, and may beadministered over longer periods of time. Dosage regimens can includetid, bid, qd, qod, biw, tiw, qw, qow, three times per month, or monthlyadministrations. Some embodiments provide any of the above-describedmethods in which the desired dosage of IFN-α is administeredsubcutaneously to the patient by bolus delivery qd, qod, tiw, biw, qw,qow, three times per month, or monthly, or is administeredsubcutaneously to the patient per day by substantially continuous orcontinuous delivery, for the desired treatment duration. In otherembodiments, any of the above-described methods may be practiced inwhich the desired dosage of PEGylated IFN-α (PEG-IFN-α) is administeredsubcutaneously to the patient by bolus delivery qw, qow, three times permonth, or monthly for the desired treatment duration.

In other embodiments, an NS3 inhibitor compound and a Type II interferonreceptor agonist are co-administered in the treatment methods of theembodiments. Type II interferon receptor agonists suitable for useherein include any interferon-γ (IFN-γ).

Effective dosages of IFN-γ can range from about 0.5 μg/m² to about 500μg/m², usually from about 1.5 μg/m² to 200 μg/m², depending on the sizeof the patient. This activity is based on 10⁶ international units (U)per 50 μg of protein. IFN-γ can be administered daily, every other day,three times a week, or substantially continuously or continuously.

In specific embodiments of interest, IFN-γ is administered to anindividual in a unit dosage form of from about 25 μg to about 500 μg,from about 50 μg to about 400 μg, or from about 100 μg to about 300 μg.In particular embodiments of interest, the dose is about 200 μg IFN-γ.In many embodiments of interest, IFN-γ1b is administered.

Where the dosage is 200 μg IFN-γ per dose, the amount of IFN-γ per bodyweight (assuming a range of body weights of from about 45 kg to about135 kg) is in the range of from about 4.4 μg IFN-γ per kg body weight toabout 1.48 μg IFN-γ per kg body weight.

The body surface area of subject individuals generally ranges from about1.33 m² to about 2.50 m². Thus, in many embodiments, an IFN-γ dosageranges from about 150 μg/m² to about 20 μg/m². For example, an IFN-γdosage ranges from about 20 μg/m² to about 30 μg/m², from about 30 μg/m²to about 40 μg/m², from about 40 μg/m² to about 50 μg/m², from about 50μg/m² to about 60 μg/m², from about 60 μg/m² to about 70 μg/m², fromabout 70 μg/m² to about 80 μg/m², from about 80 μg/m² to about 90 μg/m²,from about 90 μg/m² to about 100 μg/m², from about 100 μg/m² to about110 μg/m², from about 110 μg/m² to about 120 μg/m², from about 120 μg/m²to about 130 μg/m², from about 130 μg/m² to about 140 μg/m², or fromabout 140 μg/m² to about 150 μg/m². In some embodiments, the dosagegroups range from about 25 μg/m² to about 100 μg/m². In otherembodiments, the dosage groups range from about 25 μg/m² to about 50μg/m².

In some embodiments, a Type I or a Type III interferon receptor agonistis administered in a first dosing regimen, followed by a second dosingregimen. The first dosing regimen of Type I or a Type III interferonreceptor agonist (also referred to as “the induction regimen”) generallyinvolves administration of a higher dosage of the Type I or Type IIIinterferon receptor agonist. For example, in the case of Infergen®consensus IFN-α (CIFN), the first dosing regimen comprises administeringCIFN at about 9 μg, about 15 μg, about 18 μg, or about 27 μg. The firstdosing regimen can encompass a single dosing event, or at least two ormore dosing events. The first dosing regimen of the Type I or Type IIIinterferon receptor agonist can be administered daily, every other day,three times a week, every other week, three times per month, oncemonthly, substantially continuously or continuously.

The first dosing regimen of the Type I or Type III interferon receptoragonist is administered for a first period of time, which time periodcan be at least about 4 weeks, at least about 8 weeks, or at least about12 weeks.

The second dosing regimen of the Type I or Type III interferon receptoragonist (also referred to as “the maintenance dose”) generally involvesadministration of a lower amount of the Type I or Type III interferonreceptor agonist. For example, in the case of CIFN, the second dosingregimen comprises administering CIFN at a dose of at least about 3 μg,at least about 9 μg, at least about 15 μg, or at least about 18 μg. Thesecond dosing regimen can encompass a single dosing event, or at leasttwo or more dosing events.

The second dosing regimen of the Type I or Type III interferon receptoragonist can be administered daily, every other day, three times a week,every other week, three times per month, once monthly, substantiallycontinuously or continuously.

In some embodiments, where an “induction”/“maintenance” dosing regimenof a Type I or a Type III interferon receptor agonist is administered, a“priming” dose of a Type II interferon receptor agonist (e.g., IFN-γ) isincluded. In these embodiments, IFN-γ is administered for a period oftime from about 1 day to about 14 days, from about 2 days to about 10days, or from about 3 days to about 7 days, before the beginning oftreatment with the Type I or Type III interferon receptor agonist. Thisperiod of time is referred to as the “priming” phase.

In some of these embodiments, the Type II interferon receptor agonisttreatment is continued throughout the entire period of treatment withthe Type I or Type III interferon receptor agonist. In otherembodiments, the Type II interferon receptor agonist treatment isdiscontinued before the end of treatment with the Type I or Type IIIinterferon receptor agonist. In these embodiments, the total time oftreatment with Type II interferon receptor agonist (including the“priming” phase) is from about 2 days to about 30 days, from about 4days to about 25 days, from about 8 days to about 20 days, from about 10days to about 18 days, or from about 12 days to about 16 days. In stillother embodiments, the Type II interferon receptor agonist treatment isdiscontinued once Type I or a Type III interferon receptor agonisttreatment begins.

In other embodiments, the Type I or Type III interferon receptor agonistis administered in single dosing regimen. For example, in the case ofCIFN, the dose of CIFN is generally in a range of from about 3 μg toabout 15 μg, or from about 9 μg to about 15 μg. The dose of Type I or aType III interferon receptor agonist is generally administered daily,every other day, three times a week, every other week, three times permonth, once monthly, or substantially continuously. The dose of the TypeI or Type III interferon receptor agonist is administered for a periodof time, which period can be, for example, from at least about 24 weeksto at least about 48 weeks, or longer.

In some embodiments, where a single dosing regimen of a Type I or a TypeIII interferon receptor agonist is administered, a “priming” dose of aType II interferon receptor agonist (e.g., IFN-γ) is included. In theseembodiments, IFN-γ is administered for a period of time from about 1 dayto about 14 days, from about 2 days to about 10 days, or from about 3days to about 7 days, before the beginning of treatment with the Type Ior Type III interferon receptor agonist. This period of time is referredto as the “priming” phase. In some of these embodiments, the Type IIinterferon receptor agonist treatment is continued throughout the entireperiod of treatment with the Type I or Type III interferon receptoragonist. In other embodiments, the Type II interferon receptor agonisttreatment is discontinued before the end of treatment with the Type I orType III interferon receptor agonist. In these embodiments, the totaltime of treatment with the Type II interferon receptor agonist(including the “priming” phase) is from about 2 days to about 30 days,from about 4 days to about 25 days, from about 8 days to about 20 days,from about 10 days to about 18 days, or from about 12 days to about 16days. In still other embodiments, Type II interferon receptor agonisttreatment is discontinued once Type I or a Type III interferon receptoragonist treatment begins.

In additional embodiments, an NS3 inhibitor compound, a Type I or IIIinterferon receptor agonist, and a Type II interferon receptor agonistare co-administered for the desired duration of treatment in the methodsdescribed herein. In some embodiments, an NS3 inhibitor compound, aninterferon-α, and an interferon-γ are co-administered for the desiredduration of treatment in the methods described herein.

In some embodiments, the invention provides methods using an amount of aType I or Type III interferon receptor agonist, a Type II interferonreceptor agonist, and an NS3 inhibitor compound, effective for thetreatment of HCV infection in a patient. Some embodiments providemethods using an effective amount of an IFN-α, IFN-γ, and an NS3inhibitor compound in the treatment of HCV infection in a patient. Oneembodiment provides a method using an effective amount of a consensusIFN-α, IFN-γ and an NS3 inhibitor compound in the treatment of HCVinfection in a patient.

In general, an effective amount of a consensus interferon (CIFN) andIFN-γ suitable for use in the methods of the embodiments is provided bya dosage ratio of 1 μg CIFN:10 μg IFN-γ, where both CIFN and IFN-γ areunPEGylated and unglycosylated species.

In one embodiment, the invention provides any of the above-describedmethods modified to use an effective amount of INFERGEN® consensus IFN-αand IFN-γ in the treatment of HCV infection in a patient comprisingadministering to the patient a dosage of INFERGEN® containing an amountof about 1 μg to about 30 μg, of drug per dose of INFERGEN®,subcutaneously qd, qod, tiw, biw, qw, qow, three times per month, oncemonthly, or per day substantially continuously or continuously, incombination with a dosage of IFN-γ containing an amount of about 10 μgto about 300 μg of drug per dose of IFN-γ, subcutaneously qd, qod, tiw,biw, qw, qow, three times per month, once monthly, or per daysubstantially continuously or continuously, for the desired duration oftreatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of INFERGEN® consensus IFN-α and IFN-γ in thetreatment of virus infection in a patient comprising administering tothe patient a dosage of INFERGEN® containing an amount of about 1 μg toabout 9 μg, of drug per dose of INFERGEN®, subcutaneously qd, qod, tiw,biw, qw, qow, three times per month, once monthly, or per daysubstantially continuously or continuously, in combination with a dosageof IFN-γ containing an amount of about 10 μg to about 100 μg of drug perdose of IFN-γ, subcutaneously qd, qod, tiw, biw, qw, qow, three timesper month, once monthly, or per day substantially continuously orcontinuously, for the desired duration of treatment with an NS3inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of INFERGEN® consensus IFN-α and IFN-γ in thetreatment of virus infection in a patient comprising administering tothe patient a dosage of INFERGEN® containing an amount of about 1 μg ofdrug per dose of INFERGEN®, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or per day substantiallycontinuously or continuously, in combination with a dosage of IFN-γcontaining an amount of about 10 μg to about 50 μg of drug per dose ofIFN-γ, subcutaneously qd, qod, tiw, biw, qw, qow, three times per month,once monthly, or per day substantially continuously or continuously, forthe desired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of INFERGEN® consensus IFN-α and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of INFERGEN® containing an amount of about 9 μg ofdrug per dose of INFERGEN®, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or per day substantiallycontinuously or continuously, in combination with a dosage of IFN-γcontaining an amount of about 90 μg to about 100 μg of drug per dose ofIFN-γ, subcutaneously qd, qod, tiw, biw, qw, qow, three times per month,once monthly, or per day substantially continuously or continuously, forthe desired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of INFERGEN® consensus IFN-α and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of INFERGEN® containing an amount of about 30 μg ofdrug per dose of INFERGEN®, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or per day substantiallycontinuously or continuously, in combination with a dosage of IFN-γcontaining an amount of about 200 μg to about 300 μg of drug per dose ofIFN-γ, subcutaneously qd, qod, tiw, biw, qw, qow, three times per month,once monthly, or per day substantially continuously or continuously, forthe desired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGylated consensus IFN-α and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of PEGylated consensus IFN-α (PEG-CIFN) containingan amount of about 4 μg to about 60 μg of CIFN amino acid weight perdose of PEG-CIFN, subcutaneously qw, qow, three times per month, ormonthly, in combination with a total weekly dosage of IFN-γ containingan amount of about 30 μg to about 1,000 μg of drug per week in divideddoses administered subcutaneously qd, qod, tiw, biw, or administeredsubstantially continuously or continuously, for the desired duration oftreatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGylated consensus IFN-α and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of PEGylated consensus IFN-α (PEG-CIFN) containingan amount of about 18 μg to about 24 μg of CIFN amino acid weight perdose of PEG-CIFN, subcutaneously qw, qow, three times per month, ormonthly, in combination with a total weekly dosage of IFN-γ containingan amount of about 100 μg to about 300 μg of drug per week in divideddoses administered subcutaneously qd, qod, tiw, biw, or substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

In general, an effective amount of IFN-α2a or 2b or 2c and IFN-γsuitable for use in the methods of the embodiments is provided by adosage ratio of 1 million Units (MU) IFN-α2a or 2b or 2c: 30 μg IFN-γ,where both IFN-α2a or 2b or 2c and IFN-γ are unPEGylated andunglycosylated species.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α2a or 2b or 2c and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of IFN-α 2a, 2b or 2c containing an amount of about1 MU to about 20 MU of drug per dose of IFN-α 2a, 2b or 2csubcutaneously qd, qod, tiw, biw, or per day substantially continuouslyor continuously, in combination with a dosage of IFN-γ containing anamount of about 30 μg to about 600 μg of drug per dose of IFN-γ,subcutaneously qd, qod, tiw, biw, or per day substantially continuouslyor continuously, for the desired duration of treatment with an NS3inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α2a or 2b or 2c and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of IFN-α 2a, 2b or 2c containing an amount of about3 MU of drug per dose of IFN-α 2a, 2b or 2c subcutaneously qd, qod, tiw,biw, or per day substantially continuously or continuously, incombination with a dosage of IFN-γ containing an amount of about 100 μgof drug per dose of IFN-γ, subcutaneously qd, qod, tiw, biw, or per daysubstantially continuously or continuously, for the desired duration oftreatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α2a or 2b or 2c and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of IFN-α 2a, 2b or 2c containing an amount of about10 MU of drug per dose of IFN-α 2a, 2b or 2c subcutaneously qd, qod,tiw, biw, or per day substantially continuously or continuously, incombination with a dosage of IFN-γ containing an amount of about 300 μgof drug per dose of IFN-γ, subcutaneously qd, qod, tiw, biw, or per daysubstantially continuously or continuously, for the desired duration oftreatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGASYS®PEGylated IFN-α2a and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of PEGASYS® containing an amount of about 90 μg toabout 360 μg, of drug per dose of PEGASYS®, subcutaneously qw, qow,three times per month, or monthly, in combination with a total weeklydosage of IFN-γ containing an amount of about 30 μg to about 1,000 μg,of drug per week administered in divided doses subcutaneously qd, qod,tiw, or biw, or administered substantially continuously or continuously,for the desired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGASYS®PEGylated IFN-α2a and IFN-γ in thetreatment of a virus infection in a patient comprising administering tothe patient a dosage of PEGASYS® containing an amount of about 180 μg ofdrug per dose of PEGASYS®, subcutaneously qw, qow, three times permonth, or monthly, in combination with a total weekly dosage of IFN-γcontaining an amount of about 100 μg to about 300 μg, of drug per weekadministered in divided doses subcutaneously qd, qod, tiw, or biw, oradministered substantially continuously or continuously, for the desiredduration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEG-INTRON®PEGylated IFN-α2b and IFN-γ inthe treatment of a virus infection in a patient comprising administeringto the patient a dosage of PEG-INTRON® containing an amount of about0.75 μg to about 3.0 μg of drug per kilogram of body weight per dose ofPEG-INTRON®, subcutaneously qw, qow, three times per month, or monthly,in combination with a total weekly dosage of IFN-γ containing an amountof about 30 μg to about 1,000 μg of drug per week administered individed doses subcutaneously qd, qod, tiw, or biw, or administeredsubstantially continuously or continuously, for the desired duration oftreatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEG-INTRON®PEGylated IFN-α2b and IFN-γ inthe treatment of a virus infection in a patient comprising administeringto the patient a dosage of PEG-INTRON® containing an amount of about 1.5μg of drug per kilogram of body weight per dose of PEG-INTRON®,subcutaneously qw, qow, three times per month, or monthly, incombination with a total weekly dosage of IFN-γ containing an amount ofabout 100 μg to about 300 μg of drug per week administered in divideddoses subcutaneously qd, qod, tiw, or biw, or administered substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw, and ribavirinadministered orally qd, where the duration of therapy is 48 weeks. Inthis embodiment, ribavirin is administered in an amount of 1000 mg forindividuals weighing less than 75 kg, and 1200 mg for individualsweighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; 50 μg Actimmune®human IFN-γ1b administered subcutaneously tiw; and ribavirinadministered orally qd, where the duration of therapy is 48 weeks. Inthis embodiment, ribavirin is administered in an amount of 1000 mg forindividuals weighing less than 75 kg, and 1200 mg for individualsweighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; 100 μg Actimmune®human IFN-γ1b administered subcutaneously tiw; and ribavirinadministered orally qd, where the duration of therapy is 48 weeks. Inthis embodiment, ribavirin is administered in an amount of 1000 mg forindividuals weighing less than 75 kg, and 1200 mg for individualsweighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; and 50 μgActimmune® human IFN-γ1b administered subcutaneously tiw, where theduration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; and 100 μgActimmune® human IFN-γ1b administered subcutaneously tiw, where theduration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; 25 μg Actimmune®human IFN-γ1b administered subcutaneously tiw; and ribavirinadministered orally qd, where the duration of therapy is 48 weeks. Inthis embodiment, ribavirin is administered in an amount of 1000 mg forindividuals weighing less than 75 kg, and 1200 mg for individualsweighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; 200 μg Actimmune®human IFN-γ1b administered subcutaneously tiw; and ribavirinadministered orally qd, where the duration of therapy is 48 weeks. Inthis embodiment, ribavirin is administered in an amount of 1000 mg forindividuals weighing less than 75 kg, and 1200 mg for individualsweighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; and 25 μgActimmune® human IFN-γ1b administered subcutaneously tiw, where theduration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 9 μg INFERGEN®consensus IFN-α administered subcutaneously qd or tiw; and 200 μgActimmune® human IFN-γ1b administered subcutaneously tiw, where theduration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 100 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw, and ribavirin administered orally qd, where the durationof therapy is 48 weeks. In this embodiment, ribavirin is administered inan amount of 1000 mg for individuals weighing less than 75 kg, and 1200mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 100 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 100 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 100 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 100 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 150 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw, and ribavirin administered orally qd, where the durationof therapy is 48 weeks. In this embodiment, ribavirin is administered inan amount of 1000 mg for individuals weighing less than 75 kg, and 1200mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 150 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 150 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 150 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 150 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 200 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw, and ribavirin administered orally qd, where the durationof therapy is 48 weeks. In this embodiment, ribavirin is administered inan amount of 1000 mg for individuals weighing less than 75 kg, and 1200mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 200 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 200 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw; and ribavirin administered orally qd, where theduration of therapy is 48 weeks. In this embodiment, ribavirin isadministered in an amount of 1000 mg for individuals weighing less than75 kg, and 1200 mg for individuals weighing 75 kg or more.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 200 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 50 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

One embodiment provides any of the above-described methods modified tocomprise administering to an individual having an HCV infection aneffective amount of an NS3 inhibitor; and a regimen of 200 μg monoPEG(30 kD, linear)-ylated consensus IFN-α administered subcutaneously every10 days or qw; and 100 μg Actimmune® human IFN-γ1b administeredsubcutaneously tiw, where the duration of therapy is 48 weeks.

Any of the above-described methods involving administering an NS3inhibitor, a Type I interferon receptor agonist (e.g., an IFN-α), and aType II interferon receptor agonist (e.g., an IFN-γ), can be augmentedby administration of an effective amount of a TNF-α antagonist (e.g., aTNF-α antagonist other than pirfenidone or a pirfenidone analog).Exemplary, non-limiting TNF-α antagonists that are suitable for use insuch combination therapies include ENBREL®, REMICADE®, and HUMIRA™.

One embodiment provides a method using an effective amount of ENBREL®;an effective amount of IFN-α; an effective amount of IFN-γ; and aneffective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageENBREL® containing an amount of from about 0.1 μg to about 23 mg perdose, from about 0.1 μg to about 1 μg, from about 1 μg to about 10 μg,from about 10 μg to about 100 μg, from about 100 μg to about 1 mg, fromabout 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10mg to about 15 mg, from about 15 mg to about 20 mg, or from about 20 mgto about 23 mg of ENBREL®, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or once every other month, or perday substantially continuously or continuously, for the desired durationof treatment.

One embodiment provides a method using an effective amount of REMICADE®,an effective amount of IFN-α; an effective amount of IFN-γ; and aneffective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageof REMICADE® containing an amount of from about 0.1 mg/kg to about 4.5mg/kg, from about 0.1 mg/kg to about 0.5 mg/kg, from about 0.5 mg/kg toabout 1.0 mg/kg, from about 1.0 mg/kg to about 1.5 mg/kg, from about 1.5mg/kg to about 2.0 mg/kg, from about 2.0 mg/kg to about 2.5 mg/kg, fromabout 2.5 mg/kg to about 3.0 mg/kg, from about 3.0 mg/kg to about 3.5mg/kg, from about 3.5 mg/kg to about 4.0 mg/kg, or from about 4.0 mg/kgto about 4.5 mg/kg per dose of REMICADE®, intravenously qd, qod, tiw,biw, qw, qow, three times per month, once monthly, or once every othermonth, or per day substantially continuously or continuously, for thedesired duration of treatment.

One embodiment provides a method using an effective amount of HUMIRA™,an effective amount of IFN-α; an effective amount of IFN-γ; and aneffective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageof HUMIRA™ containing an amount of from about 0.1 μg to about 35 mg,from about 0.1 μg to about 1 μg, from about 1 μg to about 10 μg, fromabout 10 μg to about 100 μg, from about 100 μg to about 1 mg, from about1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg toabout 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about25 mg, from about 25 mg to about 30 mg, or from about 30 mg to about 35mg per dose of a HUMIRA™, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or once every other month, or perday substantially continuously or continuously, for the desired durationof treatment.

Combination Therapies with Pirfenidone

In many embodiments, the methods provide for combination therapycomprising administering an NS3 inhibitor compound as described above,and an effective amount of pirfenidone or a pirfenidone analog. In someembodiments, an NS3 inhibitor compound, one or more interferon receptoragonist(s), and pirfenidone or pirfenidone analog are co-administered inthe treatment methods of the embodiments. In certain embodiments, an NS3inhibitor compound, a Type I interferon receptor agonist, andpirfenidone (or a pirfenidone analog) are co-administered. In otherembodiments, an NS3 inhibitor compound, a Type I interferon receptoragonist, a Type II interferon receptor agonist, and pirfenidone (or apirfenidone analog) are co-administered. Type I interferon receptoragonists suitable for use herein include any IFN-α, such as interferonalfa-2a, interferon alfa-2b, interferon alfacon-1, and PEGylatedIFN-α's, such as peginterferon alfa-2a, peginterferon alfa-2b, andPEGylated consensus interferons, such as monoPEG (30 kD, linear)-ylatedconsensus interferon. Type II interferon receptor agonists suitable foruse herein include any interferon-γ.

Pirfenidone or a pirfenidone analog can be administered once per month,twice per month, three times per month, once per week, twice per week,three times per week, four times per week, five times per week, sixtimes per week, daily, or in divided daily doses ranging from once dailyto 5 times daily over a period of time ranging from about one day toabout one week, from about two weeks to about four weeks, from about onemonth to about two months, from about two months to about four months,from about four months to about six months, from about six months toabout eight months, from about eight months to about 1 year, from about1 year to about 2 years, or from about 2 years to about 4 years, ormore.

Effective dosages of pirfenidone or a specific pirfenidone analoginclude a weight-based dosage in the range from about 5 mg/kg/day toabout 125 mg/kg/day, or a fixed dosage of about 400 mg to about 3600 mgper day, or about 800 mg to about 2400 mg per day, or about 1000 mg toabout 1800 mg per day, or about 1200 mg to about 1600 mg per day,administered orally in one to five divided doses per day. Other dosesand formulations of pirfenidone and specific pirfenidone analogssuitable for use in the treatment of fibrotic diseases are described inU.S. Pat. Nos. 5,310,562; 5,518,729; 5,716,632; and 6,090,822.

One embodiment provides any of the above-described methods modified toinclude co-administering to the patient a therapeutically effectiveamount of pirfenidone or a pirfenidone analog for the duration of thedesired course of NS3 inhibitor compound treatment.

Combination Therapies with TNF-α Antagonists

In many embodiments, the methods provide for combination therapycomprising administering an effective amount of an NS3 inhibitorcompound as described above, and an effective amount of TNF-αantagonist, in combination therapy for treatment of an HCV infection.

Effective dosages of a TNF-α antagonist range from 0.1 μg to 40 mg perdose, e.g., from about 0.1 μg to about 0.5 μg per dose, from about 0.5μg to about 1.0 μg per dose, from about 1.0 μg per dose to about 5.0 μgper dose, from about 5.0 μg to about 10 μg per dose, from about 10 μg toabout 20 μg per dose, from about 20 μg per dose to about 30 μg per dose,from about 30 μg per dose to about 40 μg per dose, from about 40 μg perdose to about 50 μg per dose, from about 50 μg per dose to about 60 μgper dose, from about 60 μg per dose to about 70 μg per dose, from about70 μg to about 80 μg per dose, from about 80 μg per dose to about 100 μgper dose, from about 100 μg to about 150 μg per dose, from about 150 μgto about 200 μg per dose, from about 200 μg per dose to about 250 μg perdose, from about 250 μg to about 300 μg per dose, from about 300 μg toabout 400 μg per dose, from about 400 μg to about 500 μg per dose, fromabout 500 μg to about 600 μg per dose, from about 600 μg to about 700 μgper dose, from about 700 μg to about 800 μg per dose, from about 800 μgto about 900 μg per dose, from about 900 μg to about 1000 μg per dose,from about 1 mg to about 10 mg per dose, from about 10 mg to about 15 mgper dose, from about 15 mg to about 20 mg per dose, from about 20 mg toabout 25 mg per dose, from about 25 mg to about 30 mg per dose, fromabout 30 mg to about 35 mg per dose, or from about 35 mg to about 40 mgper dose.

In some embodiments, effective dosages of a TNF-α antagonist areexpressed as mg/kg body weight. In these embodiments, effective dosagesof a TNF-α antagonist are from about 0.1 mg/kg body weight to about 10mg/kg body weight, e.g., from about 0.1 mg/kg body weight to about 0.5mg/kg body weight, from about 0.5 mg/kg body weight to about 1.0 mg/kgbody weight, from about 1.0 mg/kg body weight to about 2.5 mg/kg bodyweight, from about 2.5 mg/kg body weight to about 5.0 mg/kg body weight,from about 5.0 mg/kg body weight to about 7.5 mg/kg body weight, or fromabout 7.5 mg/kg body weight to about 10 mg/kg body weight.

In many embodiments, a TNF-α antagonist is administered for a period ofabout 1 day to about 7 days, or about 1 week to about 2 weeks, or about2 weeks to about 3 weeks, or about 3 weeks to about 4 weeks, or about 1month to about 2 months, or about 3 months to about 4 months, or about 4months to about 6 months, or about 6 months to about 8 months, or about8 months to about 12 months, or at least one year, and may beadministered over longer periods of time. The TNF-α antagonist can beadministered tid, bid, qd, qod, biw, tiw, qw, qow, three times permonth, once monthly, substantially continuously, or continuously.

In many embodiments, multiple doses of a TNF-α antagonist areadministered. For example, a TNF-α antagonist is administered once permonth, twice per month, three times per month, every other week (qow),once per week (qw), twice per week (biw), three times per week (tiw),four times per week, five times per week, six times per week, everyother day (qod), daily (qd), twice a day (bid), or three times a day(tid), substantially continuously, or continuously, over a period oftime ranging from about one day to about one week, from about two weeksto about four weeks, from about one month to about two months, fromabout two months to about four months, from about four months to aboutsix months, from about six months to about eight months, from abouteight months to about 1 year, from about 1 year to about 2 years, orfrom about 2 years to about 4 years, or more.

A TNF-α antagonist and an NS3 inhibitor are generally administered inseparate formulations. A TNF-α antagonist and an NS3 inhibitor may beadministered substantially simultaneously, or within about 30 minutes,about 1 hour, about 2 hours, about 4 hours, about 8 hours, about 16hours, about 24 hours, about 36 hours, about 72 hours, about 4 days,about 7 days, or about 2 weeks of one another.

One embodiment provides a method using an effective amount of a TNF-αantagonist and an effective amount of an NS3 inhibitor in the treatmentof an HCV infection in a patient, comprising administering to thepatient a dosage of a TNF-α antagonist containing an amount of fromabout 0.1 μg to about 40 mg per dose of a TNF-α antagonist,subcutaneously qd, qod, tiw, or biw, or per day substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

One embodiment provides a method using an effective amount of ENBREL®and an effective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageENBREL® containing an amount of from about 0.1 μg to about 23 mg perdose, from about 0.1 μg to about 1 μg, from about 1 μg to about 10 μg,from about 10 μg to about 100 μg, from about 100 μg to about 1 mg, fromabout 1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10mg to about 15 mg, from about 15 mg to about 20 mg, or from about 20 mgto about 23 mg of ENBREL®, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or once every other month, or perday substantially continuously or continuously, for the desired durationof treatment with an NS3 inhibitor compound.

One embodiment provides a method using an effective amount of REMICADE®and an effective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageof REMICADE® containing an amount of from about 0.1 mg/kg to about 4.5mg/kg, from about 0.1 mg/kg to about 0.5 mg/kg, from about 0.5 mg/kg toabout 1.0 mg/kg, from about 1.0 mg/kg to about 1.5 mg/kg, from about 1.5mg/kg to about 2.0 mg/kg, from about 2.0 mg/kg to about 2.5 mg/kg, fromabout 2.5 mg/kg to about 3.0 mg/kg, from about 3.0 mg/kg to about 3.5mg/kg, from about 3.5 mg/kg to about 4.0 mg/kg, or from about 4.0 mg/kgto about 4.5 mg/kg per dose of REMICADE®, intravenously qd, qod, tiw,biw, qw, qow, three times per month, once monthly, or once every othermonth, or per day substantially continuously or continuously, for thedesired duration of treatment with an NS3 inhibitor compound.

One embodiment provides a method using an effective amount of HUMIRA™and an effective amount of an NS3 inhibitor in the treatment of an HCVinfection in a patient, comprising administering to the patient a dosageof HUMIRA™ containing an amount of from about 0.1 μg to about 35 mg,from about 0.1 μg to about 1 μg, from about 1 μg to about 10 μg, fromabout 10 μg to about 100 μg, from about 100 μg to about 1 mg, from about1 mg to about 5 mg, from about 5 mg to about 10 mg, from about 10 mg toabout 15 mg, from about 15 mg to about 20 mg, from about 20 mg to about25 mg, from about 25 mg to about 30 mg, or from about 30 mg to about 35mg per dose of a HUMIRA™, subcutaneously qd, qod, tiw, biw, qw, qow,three times per month, once monthly, or once every other month, or perday substantially continuously or continuously, for the desired durationof treatment with an NS3 inhibitor compound.

Combination Therapies with Thymosin-α

In many embodiments, the methods provide for combination therapycomprising administering an effective amount of an NS3 inhibitorcompound as described above, and an effective amount of thymosin-α, incombination therapy for treatment of an HCV infection.

Effective dosages of thymosin-α range from about 0.5 mg to about 5 mg,e.g., from about 0.5 mg to about 1.0 mg, from about 1.0 mg to about 1.5mg, from about 1.5 mg to about 2.0 mg, from about 2.0 mg to about 2.5mg, from about 2.5 mg to about 3.0 mg, from about 3.0 mg to about 3.5mg, from about 3.5 mg to about 4.0 mg, from about 4.0 mg to about 4.5mg, or from about 4.5 mg to about 5.0 mg. In particular embodiments,thymosin-α is administered in dosages containing an amount of 1.0 mg or1.6 mg.

One embodiment provides a method using an effective amount of ZADAXIN™thymosin-α and an effective amount of an NS3 inhibitor in the treatmentof an HCV infection in a patient, comprising administering to thepatient a dosage of ZADAXIN™ containing an amount of from about 1.0 mgto about 1.6 mg per dose, subcutaneously twice per week for the desiredduration of treatment with the NS3 inhibitor compound.

Combination Therapies with a TNF-α Antagonist and an Interferon

Some embodiments provide a method of treating an HCV infection in anindividual having an HCV infection, the method comprising administeringan effective amount of an NS3 inhibitor, and effective amount of a TNF-αantagonist, and an effective amount of one or more interferons.

One embodiment provides any of the above-described methods modified touse an effective amount of IFN-γ and an effective amount of a TNF-αantagonist in the treatment of HCV infection in a patient comprisingadministering to the patient a dosage of IFN-γ containing an amount ofabout 10 μg to about 300 μg of drug per dose of IFN-γ, subcutaneouslyqd, qod, tiw, biw, qw, qow, three times per month, once monthly, or perday substantially continuously or continuously, in combination with adosage of a TNF-α antagonist containing an amount of from about 0.1 μgto about 40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod,tiw, or biw, or per day substantially continuously or continuously, forthe desired duration of treatment with an NS3 inhibitor compound.

One embodiment provides any of the above-described methods modified touse an effective amount of IFN-γ and an effective amount of a TNF-αantagonist in the treatment of HCV infection in a patient comprisingadministering to the patient a dosage of IFN-γ containing an amount ofabout 10 μg to about 100 μg of drug per dose of IFN-γ, subcutaneouslyqd, qod, tiw, biw, qw, qow, three times per month, once monthly, or perday substantially continuously or continuously, in combination with adosage of a TNF-α antagonist containing an amount of from about 0.1 μgto about 40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod,tiw, or biw, or per day substantially continuously or continuously, forthe desired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-γ and an effective amount of a TNF-αantagonist in the treatment of a virus infection in a patient comprisingadministering to the patient a total weekly dosage of IFN-γ containingan amount of about 30 μg to about 1,000 μg of drug per week in divideddoses administered subcutaneously qd, qod, tiw, biw, or administeredsubstantially continuously or continuously, in combination with a dosageof a TNF-α antagonist containing an amount of from about 0.1 μg to about40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, orbiw, or per day substantially continuously or continuously, for thedesired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-γ and an effective amount of a TNF-αantagonist in the treatment of a virus infection in a patient comprisingadministering to the patient a total weekly dosage of IFN-γ containingan amount of about 100 μg to about 300 μg of drug per week in divideddoses administered subcutaneously qd, qod, tiw, biw, or administeredsubstantially continuously or continuously, in combination with a dosageof a TNF-α antagonist containing an amount of from about 0.1 μg to about40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, orbiw, or per day substantially continuously or continuously, for thedesired duration of treatment with an NS3 inhibitor compound.

One embodiment provides any of the above-described methods modified touse an effective amount of INFERGEN® consensus IFN-α and a TNF-αantagonist in the treatment of HCV infection in a patient comprisingadministering to the patient a dosage of INFERGEN® containing an amountof about 1 μg to about 30 μg, of drug per dose of INFERGEN®,subcutaneously qd, qod, tiw, biw, qw, qow, three times per month, oncemonthly, or per day substantially continuously or continuously, incombination with a dosage of a TNF-α antagonist containing an amount offrom about 0.1 μg to about 40 mg per dose of a TNF-α antagonist,subcutaneously qd, qod, tiw, or biw, or per day substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

One embodiment provides any of the above-described methods modified touse an effective amount of INFERGEN® consensus IFN-α and a TNF-αantagonist in the treatment of HCV infection in a patient comprisingadministering to the patient a dosage of INFERGEN® containing an amountof about 1 μg to about 9 μg, of drug per dose of INFERGEN®,subcutaneously qd, qod, tiw, biw, qw, qow, three times per month, oncemonthly, or per day substantially continuously or continuously, incombination with a dosage of a TNF-α antagonist containing an amount offrom about 0.1 μg to about 40 mg per dose of a TNF-α antagonist,subcutaneously qd, qod, tiw, or biw, or per day substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGylated consensus IFN-α and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of PEGylatedconsensus IFN-α (PEG-CIFN) containing an amount of about 4 μg to about60 μg of CIFN amino acid weight per dose of PEG-CIFN, subcutaneously qw,qow, three times per month, or monthly, in combination with a dosage ofa TNF-α antagonist containing an amount of from about 0.1 μg to about 40mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, or biw,or per day substantially continuously or continuously, for the desiredduration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGylated consensus IFN-α and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of PEGylatedconsensus IFN-α (PEG-CIFN) containing an amount of about 18 μg to about24 μg of CIFN amino acid weight per dose of PEG-CIFN, subcutaneously qw,qow, three times per month, or monthly, in combination with a dosage ofa TNF-α antagonist containing an amount of from about 0.1 μg to about 40mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, or biw,or per day substantially continuously or continuously, for the desiredduration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α2a or 2b or 2c and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of IFN-α 2a, 2bor 2c containing an amount of about 1 MU to about 20 MU of drug per doseof IFN-α 2a, 2b or 2c subcutaneously qd, qod, tiw, biw, or per daysubstantially continuously or continuously, in combination with a dosageof a TNF-α antagonist containing an amount of from about 0.1 μg to about40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, orbiw, or per day substantially continuously or continuously, for thedesired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α 2a or 2b or 2c and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of IFN-α 2a, 2bor 2c containing an amount of about 3 MU of drug per dose of IFN-α 2a,2b or 2c subcutaneously qd, qod, tiw, biw, or per day substantiallycontinuously or continuously, in combination with a dosage of a TNF-αantagonist containing an amount of from about 0.1 μg to about 40 mg perdose of a TNF-α antagonist, subcutaneously qd, qod, tiw, or biw, or perday substantially continuously or continuously, for the desired durationof treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of IFN-α2a or 2b or 2c and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of IFN-α 2a, 2bor 2c containing an amount of about 10 MU of drug per dose of IFN-α 2a,2b or 2c subcutaneously qd, qod, tiw, biw, or per day substantiallycontinuously or continuously, in combination with a dosage of a TNF-αantagonist containing an amount of from about 0.1 μg to about 40 mg perdose of a TNF-α antagonist, subcutaneously qd, qod, tiw, or biw, or perday substantially continuously or continuously, for the desired durationof treatment with an NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGASYS®PEGylated IFN-α2a and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of PEGASYS®containing an amount of about 90 μg to about 360 μg, of drug per dose ofPEGASYS®, subcutaneously qw, qow, three times per month, or monthly, incombination with a dosage of a TNF-α antagonist containing an amount offrom about 0.1 μg to about 40 mg per dose of a TNF-α antagonist,subcutaneously qd, qod, tiw, or biw, or per day substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEGASYS®PEGylated IFN-α2a and an effectiveamount of a TNF-α antagonist in the treatment of a virus infection in apatient comprising administering to the patient a dosage of PEGASYS®containing an amount of about 180 μg, of drug per dose of PEGASYS®,subcutaneously qw, qow, three times per month, or monthly, incombination with a dosage of a TNF-α antagonist containing an amount offrom about 0.1 μg to about 40 mg per dose of a TNF-α antagonist,subcutaneously qd, qod, tiw, or biw, or per day substantiallycontinuously or continuously, for the desired duration of treatment withan NS3 inhibitor compound.

Another embodiment provides any of the above-described methods modifiedto use an effective amount of PEG-INTRON®PEGylated IFN-α2b and aneffective amount of a TNF-α antagonist in the treatment of a virusinfection in a patient comprising administering to the patient a dosageof PEG-INTRON® containing an amount of about 0.75 μg to about 3.0 μg ofdrug per kilogram of body weight per dose of PEG-INTRON®, subcutaneouslyqw, qow, three times per month, or monthly, in combination with a dosageof a TNF-α antagonist containing an amount of from about 0.1 μg to about40 mg per dose of a TNF-α antagonist, subcutaneously qd, qod, tiw, orbiw, or per day substantially continuously or continuously, for thedesired duration of treatment with an NS3 inhibitor compound.

Another embodiment provides any Of the above-described methods modifiedto use an effective amount of PEG-INTRON®PEGylated IFN-α2b and aneffective amount of a TNF-α antagonist in the treatment of a virusinfection in a patient comprising administering to the patient a dosageof PEG-INTRON® containing an amount of about 1.5 μg of drug per kilogramof body weight per dose of PEG-INTRON®, subcutaneously qw, qow, threetimes per month, or monthly, in combination with a dosage of a TNF-αantagonist containing an amount of from about 0.1 μg to about 40 mg perdose of a TNF-α antagonist, subcutaneously qd, qod, tiw, or biw, or perday substantially continuously or continuously, for the desired durationof treatment with an NS3 inhibitor compound.

Combination Therapies with Other Antiviral Agents

Other agents such as inhibitors of HCV NS3 helicase are also attractivedrugs for combinational therapy, and are contemplated for use incombination therapies described herein. Ribozymes such as Heptazyme™ andphosphorothioate oligonucleotides which are complementary to HCV proteinsequences and which inhibit the expression of viral core proteins arealso suitable for use in combination therapies described herein.

In some embodiments, the additional antiviral agent(s) is administeredduring the entire course of treatment with the NS3 inhibitor compounddescribed herein, and the beginning and end of the treatment periodscoincide. In other embodiments, the additional antiviral agent(s) isadministered for a period of time that is overlapping with that of theNS3 inhibitor compound treatment, e.g., treatment with the additionalantiviral agent(s) begins before the NS3 inhibitor compound treatmentbegins and ends before the NS3 inhibitor compound treatment ends;treatment with the additional antiviral agent(s) begins after the NS3inhibitor compound treatment begins and ends after the NS3 inhibitorcompound treatment ends; treatment with the additional antiviralagent(s) begins after the NS3 inhibitor compound treatment begins andends before the NS3 inhibitor compound treatment ends; or treatment withthe additional antiviral agent(s) begins before the NS3 inhibitorcompound treatment begins and ends after the NS3 inhibitor compoundtreatment ends.

The NS3 inhibitor compound can be administered together with (i.e.,simultaneously in separate formulations; simultaneously in the sameformulation; administered in separate formulations and within about 48hours, within about 36 hours, within about 24 hours, within about 16hours, within about 12 hours, within about 8 hours, within about 4hours, within about 2 hours, within about 1 hour, within about 30minutes, or within about 15 minutes or less) one or more additionalantiviral agents.

As non-limiting examples, any of the above-described methods featuringan IFN-α regimen can be modified to replace the subject IFN-α regimenwith a regimen of monoPEG (30 kD, linear)-ylated consensus IFN-αcomprising administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 100 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α regimen can be modified to replace the subject IFN-α regimenwith a regimen of monoPEG (30 kD, linear)-ylated consensus IFN-αcomprising administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 150 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α regimen can be modified to replace the subject IFN-α regimenwith a regimen of monoPEG (30 kD, linear)-ylated consensus IFN-αcomprising administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 200 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α regimen can be modified to replace the subject IFN-α regimenwith a regimen of INFERGEN® interferon alfacon-1 comprisingadministering a dosage of INFERGEN® interferon alfacon-1 containing anamount of 9 μg of drug per dose, subcutaneously once daily or threetimes per week for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α regimen can be modified to replace the subject IFN-α regimenwith a regimen of INFERGEN® interferon alfacon-1 comprisingadministering a dosage of INFERGEN® interferon alfacon-1 containing anamount of 15 μg of drug per dose, subcutaneously once daily or threetimes per week for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ regimen can be modified to replace the subject IFN-γ regimenwith a regimen of IFN-γ comprising administering a dosage of IFN-γcontaining an amount of 25 μg of drug per dose, subcutaneously threetimes per week for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ regimen can be modified to replace the subject IFN-γ regimenwith a regimen of IFN-γ comprising administering a dosage of IFN-γcontaining an amount of 50 μg of drug per dose, subcutaneously threetimes per week for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ regimen can be modified to replace the subject IFN-γ regimenwith a regimen of IFN-γ comprising administering a dosage of IFN-γcontaining an amount of 100 μg of drug per dose, subcutaneously threetimes per week for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 100 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 50 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuring aTNF antagonist regimen can be modified to replace the subject TNFantagonist regimen with a TNF antagonist regimen comprisingadministering a dosage of a TNF antagonist selected from the group of:(a) etanercept in an amount of 25 mg of drug per dose subcutaneouslytwice per week, (b) infliximab in an amount of 3 mg of drug per kilogramof body weight per dose intravenously at weeks 0, 2 and 6, and every 8weeks thereafter, or (c) adalimumab in an amount of 40 mg of drug perdose subcutaneously once weekly or once every 2 weeks; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 100 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 100 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 150 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 50 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 150 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 100 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 200 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 50 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of monoPEG(30 kD, linear)-ylated consensus IFN-α containing an amount of 200 μg ofdrug per dose, subcutaneously once weekly, once every 8 days, or onceevery 10 days; and (b) administering a dosage of IFN-γ containing anamount of 100 μg of drug per dose, subcutaneously three times per week;for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 25 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 50 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 100 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 25 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 50 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 9 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 100 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 25 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 50 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously three times per week; and (b) administering a dosage ofIFN-γ containing an amount of 100 μg of drug per dose, subcutaneouslythree times per week; for the desired treatment duration with an NS3inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 25 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 50 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and IFN-γ combination regimen can be modified to replace thesubject IFN-α and IFN-γ combination regimen with an IFN-α and IFN-γcombination regimen comprising: (a) administering a dosage of INFERGEN®interferon alfacon-1 containing an amount of 15 μg of drug per dose,subcutaneously once daily; and (b) administering a dosage of IFN-γcontaining an amount of 100 μg of drug per dose, subcutaneously threetimes per week; for the desired treatment duration with an NS3 inhibitorcompound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 100 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 100 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 100 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 50 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 150 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 50 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 150 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 100 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 200 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 50 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of monoPEG (30 kD, linear)-ylatedconsensus IFN-α containing an amount of 200 μg of drug per dose,subcutaneously once weekly, once every 8 days, or once every 10 days;(b) administering a dosage of IFN-γ containing an amount of 100 μg ofdrug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 25 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 50 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 100 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 25 μgof drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 50 μgof drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 9 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 100μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 25 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 50 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously threetimes per week; (b) administering a dosage of IFN-γ containing an amountof 100 μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 25 μgof drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 50 μgof drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α, IFN-γ and TNF antagonist combination regimen can be modifiedto replace the subject IFN-α, IFN-γ and TNF antagonist combinationregimen with an IFN-α, IFN-γ and TNF antagonist combination regimencomprising: (a) administering a dosage of INFERGEN® interferon alfacon-1containing an amount of 15 μg of drug per dose, subcutaneously oncedaily; (b) administering a dosage of IFN-γ containing an amount of 100μg of drug per dose, subcutaneously three times per week; and (c)administering a dosage of a TNF antagonist selected from (i) etanerceptin an amount of 25 mg subcutaneously twice per week, (ii) infliximab inan amount of 3 mg of drug per kilogram of body weight intravenously atweeks 0, 2 and 6, and every 8 weeks thereafter or (iii) adalimumab in anamount of 40 mg subcutaneously once weekly or once every other week; forthe desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and TNF antagonist combination regimen can be modified toreplace the subject IFN-α and TNF antagonist combination regimen with anIFN-α and TNF antagonist combination regimen comprising: (a)administering a dosage of monoPEG (30 kD, linear)-ylated consensus IFN-αcontaining an amount of 100 μg of drug per dose, subcutaneously onceweekly, once every 8 days, or once every 10 days; and (b) administeringa dosage of a TNF antagonist selected from (i) etanercept in an amountof 25 mg subcutaneously twice per week, (ii) infliximab in an amount of3 mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and TNF antagonist combination regimen can be modified toreplace the subject IFN-α and TNF antagonist combination regimen with anIFN-α and TNF antagonist combination regimen comprising: (a)administering a dosage of monoPEG (30 kD, linear)-ylated consensus IFN-αcontaining an amount of 150 μg of drug per dose, subcutaneously onceweekly, once every 8 days, or once every 10 days; and (b) administeringa dosage of a TNF antagonist selected from (i) etanercept in an amountof 25 mg subcutaneously twice per week, (ii) infliximab in an amount of3 mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and TNF antagonist combination regimen can be modified toreplace the subject IFN-α and TNF antagonist combination regimen with anIFN-α and TNF antagonist combination regimen comprising: (a)administering a dosage of monoPEG (30 kD, linear)-ylated consensus IFN-αcontaining an amount of 200 μg of drug per dose, subcutaneously onceweekly, once every 8 days, or once every 10 days; and (b) administeringa dosage of a TNF antagonist selected from (i) etanercept in an amountof 25 mg subcutaneously twice per week, (ii) infliximab in an amount of3 mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and TNF antagonist combination regimen can be modified toreplace the subject IFN-α and TNF antagonist combination regimen with anIFN-α and TNF antagonist combination regimen comprising: (a)administering a dosage of INFERGEN® interferon alfacon-1 containing anamount of 9 μg of drug per dose, subcutaneously once daily or threetimes per week; and (b) administering a dosage of a TNF antagonistselected from (i) etanercept in an amount of 25 mg subcutaneously twiceper week, (ii) infliximab in an amount of 3 mg of drug per kilogram ofbody weight intravenously at weeks 0, 2 and 6, and every 8 weeksthereafter or (iii) adalimumab in an amount of 40 mg subcutaneously onceweekly or once every other week; for the desired treatment duration withan NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-α and TNF antagonist combination regimen can be modified toreplace the subject IFN-α and TNF antagonist combination regimen with anIFN-α and TNF antagonist combination regimen comprising: (a)administering a dosage of INFERGEN® interferon alfacon-1 containing anamount of 15 μg of drug per dose, subcutaneously once daily or threetimes per week; and (b) administering a dosage of a TNF antagonistselected from (i) etanercept in an amount of 25 mg subcutaneously twiceper week, (ii) infliximab in an amount of 3 mg of drug per kilogram ofbody weight intravenously at weeks 0, 2 and 6, and every 8 weeksthereafter or (iii) adalimumab in an amount of 40 mg subcutaneously onceweekly or once every other week; for the desired treatment duration withan NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ and TNF antagonist combination regimen can be modified toreplace the subject IFN-γ and TNF antagonist combination regimen with anIFN-γ and TNF antagonist combination regimen comprising: (a)administering a dosage of IFN-γ containing an amount of 25 μg of drugper dose, subcutaneously three times per week; and (b) administering adosage of a TNF antagonist selected from (i) etanercept in an amount of25 mg subcutaneously twice per week, (ii) infliximab in an amount of 3mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ and TNF antagonist combination regimen can be modified toreplace the subject IFN-γ and TNF antagonist combination regimen with anIFN-γ and TNF antagonist combination regimen comprising: (a)administering a dosage of IFN-γ containing an amount of 50 μg of drugper dose, subcutaneously three times per week; and (b) administering adosage of a TNF antagonist selected from (i) etanercept in an amount of25 mg subcutaneously twice per week, (ii) infliximab in an amount of 3mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan IFN-γ and TNF antagonist combination regimen can be modified toreplace the subject IFN-γ and TNF antagonist combination regimen with anIFN-γ and TNF antagonist combination regimen comprising: (a)administering a dosage of IFN-γ containing an amount of 100 μg of drugper dose, subcutaneously three times per week; and (b) administering adosage of a TNF antagonist selected from (i) etanercept in an amount of25 mg subcutaneously twice per week, (ii) infliximab in an amount of 3mg of drug per kilogram of body weight intravenously at weeks 0, 2 and6, and every 8 weeks thereafter or (iii) adalimumab in an amount of 40mg subcutaneously once weekly or once every other week; for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods thatincludes a regimen of monoPEG (30 kD, linear)-ylated consensus IFN-α canbe modified to replace the regimen of monoPEG (30 kD, linear)-ylatedconsensus IFN-α with a regimen of peginterferon alfa-2a comprisingadministering a dosage of peginterferon alfa-2a containing an amount of180 μg of drug per dose, subcutaneously once weekly for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods thatincludes a regimen of monoPEG (30 kD, linear)-ylated consensus IFN-α canbe modified to replace the regimen of monoPEG (30 kD, linear)-ylatedconsensus IFN-α with a regimen of peginterferon alfa-2b comprisingadministering a dosage of peginterferon alfa-2b containing an amount of1.0 μg to 1.5 μg of drug per kilogram of body weight per dose,subcutaneously once or twice weekly for the desired treatment durationwith an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to include administering a dosage of ribavirin containing anamount of 400 mg, 800 mg, 1000 mg or 1200 mg of drug orally per day,optionally in two or more divided doses per day, for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to include administering a dosage of ribavirin containing (i)an amount of 1000 mg of drug orally per day for patients having a bodyweight of less than 75 kg or (ii) an amount of 1200 mg of drug orallyper day for patients having a body weight of greater than or equal to 75kg, optionally in two or more divided doses per day, for the desiredtreatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to replace the subject NS3 inhibitor regimen with an NS3inhibitor regimen comprising administering a dosage of 0.01 mg to 0.1 mgof drug per kilogram of body weight orally daily, optionally in two ormore divided doses per day, for the desired treatment duration with theNS3 inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to replace the subject NS3 inhibitor regimen with an NS3inhibitor regimen comprising administering a dosage of 0.1 mg to 1 mg ofdrug per kilogram of body weight orally daily, optionally in two or moredivided doses per day, for the desired treatment duration with the NS3inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to replace the subject NS3 inhibitor regimen with an NS3inhibitor regimen comprising administering a dosage of 1 mg to 10 mg ofdrug per kilogram of body weight orally daily, optionally in two or moredivided doses per day, for the desired treatment duration with the NS3inhibitor compound.

As non-limiting examples, any of the above-described methods can bemodified to replace the subject NS3 inhibitor regimen with an NS3inhibitor regimen comprising administering a dosage of 10 mg to 100 mgof drug per kilogram of body weight orally daily, optionally in two ormore divided doses per day, for the desired treatment duration with theNS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan NS5B inhibitor regimen can be modified to replace the subject NS5Binhibitor regimen with an NS5B inhibitor regimen comprisingadministering a dosage of 0.01 mg to 0.1 mg of drug per kilogram of bodyweight orally daily, optionally in two or more divided doses per day,for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan NS5B inhibitor regimen can be modified to replace the subject NS5Binhibitor regimen with an NS5B inhibitor regimen comprisingadministering a dosage of 0.1 mg to 1 mg of drug per kilogram of bodyweight orally daily, optionally in two or more divided doses per day,for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan NS5B inhibitor regimen can be modified to replace the subject NS5Binhibitor regimen with an NS5B inhibitor regimen comprisingadministering a dosage of 1 mg to 10 mg of drug per kilogram of bodyweight orally daily, optionally in two or more divided doses per day,for the desired treatment duration with an NS3 inhibitor compound.

As non-limiting examples, any of the above-described methods featuringan NS5B inhibitor regimen can be modified to replace the subject NS5Binhibitor regimen with an NS5B inhibitor regimen comprisingadministering a dosage of 10 mg to 100 mg of drug per kilogram of bodyweight orally daily, optionally in two or more divided doses per day,for the desired treatment duration with an NS3 inhibitor compound.

Patient Identification

In certain embodiments, the specific regimen of drug therapy used intreatment of the HCV patient is selected according to certain diseaseparameters exhibited by the patient, such as the initial viral load,genotype of the HCV infection in the patient, liver histology and/orstage of liver fibrosis in the patient.

Thus, some embodiments provide any of the above-described methods forthe treatment of HCV infection in which the subject method is modifiedto treat a treatment failure patient for a duration of 48 weeks.

Other embodiments provide any of the above-described methods for HCV inwhich the subject method is modified to treat a non-responder patient,where the patient receives a 48 week course of therapy.

Other embodiments provide any of the above-described methods for thetreatment of HCV infection in which the subject method is modified totreat a relapser patient, where the patient receives a 48 week course oftherapy.

Other embodiments provide any of the above-described methods for thetreatment of HCV infection in which the subject method is modified totreat a naïve patient infected with HCV genotype 1, where the patientreceives a 48 week course of therapy.

Other embodiments provide any of the above-described methods for thetreatment of HCV infection in which the subject method is modified totreat a naïve patient infected with HCV genotype 4, where the patientreceives a 48 week course of therapy.

Other embodiments provide any of the above-described methods for thetreatment of HCV infection in which the subject method is modified totreat a naïve patient infected with HCV genotype 1, where the patienthas a high viral load (HVL), where “HVL” refers to an HCV viral load ofgreater than 2×10⁶ HCV genome copies per mL serum, and where the patientreceives a 48 week course of therapy.

One embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having advanced or severestage liver fibrosis as measured by a Knodell score of 3 or 4 and then(2) administering to the patient the drug therapy of the subject methodfor a time period of about 24 weeks to about 60 weeks, or about 30 weeksto about one year, or about 36 weeks to about 50 weeks, or about 40weeks to about 48 weeks, or at least about 24 weeks, or at least about30 weeks, or at least about 36 weeks, or at least about 40 weeks, or atleast about 48 weeks, or at least about 60 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having advanced or severestage liver fibrosis as measured by a Knodell score of 3 or 4 and then(2) administering to the patient the drug therapy of the subject methodfor a time period of about 40 weeks to about 50 weeks, or about 48weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of greater than 2 million viralgenome copies per ml of patient serum and then (2) administering to thepatient the drug therapy of the subject method for a time period ofabout 24 weeks to about 60 weeks, or about 30 weeks to about one year,or about 36 weeks to about 50 weeks, or about 40 weeks to about 48weeks, or at least about 24 weeks, or at least about 30 weeks, or atleast about 36 weeks, or at least about 40 weeks, or at least about 48weeks, or at least about 60 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of greater than 2 million viralgenome copies per ml of patient serum and then (2) administering to thepatient the drug therapy of the subject method for a time period ofabout 40 weeks to about 50 weeks, or about 48 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of greater than 2 million viralgenome copies per ml of patient serum and no or early stage liverfibrosis as measured by a Knodell score of 0, 1, or 2 and then (2)administering to the patient the drug therapy of the subject method fora time period of about 24 weeks to about 60 weeks, or about 30 weeks toabout one year, or about 36 weeks to about 50 weeks, or about 40 weeksto about 48 weeks, or at least about 24 weeks, or at least about 30weeks, or at least about 36 weeks, or at least about 40 weeks, or atleast about 48 weeks, or at least about 60 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of greater than 2 million viralgenome copies per ml of patient serum and no or early stage liverfibrosis as measured by a Knodell score of 0, 1, or 2 and then (2)administering to the patient the drug therapy of the subject method fora time period of about 40 weeks to about 50 weeks, or about 48 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of less than or equal to 2 millionviral genome copies per ml of patient serum and then (2) administeringto the patient the drug therapy of the subject method for a time periodof about 20 weeks to about 50 weeks, or about 24 weeks to about 48weeks, or about 30 weeks to about 40 weeks, or up to about 20 weeks, orup to about 24 weeks, or up to about 30 Weeks, or up to about 36 weeks,or up to about 48 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of less than or equal to 2 millionviral genome copies per ml of patient serum and then (2) administeringto the patient the drug therapy of the subject method for a time periodof about 20 weeks to about 24 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1infection and an initial viral load of less than or equal to 2 millionviral genome copies per ml of patient serum and then (2) administeringto the patient the drug therapy of the subject method for a time periodof about 24 weeks to about 48 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 2or 3 infection and then (2) administering to the patient the drugtherapy of the subject method for a time period of about 24 weeks toabout 60 weeks, or about 30 weeks to about one year, or about 36 weeksto about 50 weeks, or about 40 weeks to about 48 weeks, or at leastabout 24 weeks, or at least about 30 weeks, or at least about 36 weeks,or at least about 40 weeks, or at least about 48 weeks, or at leastabout 60 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 2or 3 infection and then (2) administering to the patient the drugtherapy of the subject method for a time period of about 20 weeks toabout 50 weeks, or about 24 weeks to about 48 weeks, or about 30 weeksto about 40 weeks, or up to about 20 weeks, or up to about 24 weeks, orup to about 30 weeks, or up to about 36 weeks, or up to about 48 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 2or 3 infection and then (2) administering to the patient the drugtherapy of the subject method for a time period of about 20 weeks toabout 24 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 2or 3 infection and then (2) administering to the patient the drugtherapy of the subject method for a time period of at least about 24weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV genotype 1or 4 infection and then (2) administering to the patient the drugtherapy of the subject method for a time period of about 24 weeks toabout 60 weeks, or about 30 weeks to about one year, or about 36 weeksto about 50 weeks, or about 40 weeks to about 48 weeks, or at leastabout 24 weeks, or at least about 30 weeks, or at least about 36 weeks,or at least about 40 weeks, or at least about 48 weeks, or at leastabout 60 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV infectioncharacterized by any of HCV genotypes 5, 6, 7, 8 and 9 and then (2)administering to the patient the drug therapy of the subject method fora time period of about 20 weeks to about 50 weeks.

Another embodiment provides any of the above-described methods for thetreatment of an HCV infection, where the subject method is modified toinclude the steps of (1) identifying a patient having an HCV infectioncharacterized by any of HCV genotypes 5, 6, 7, 8 and 9 and then (2)administering to the patient the drug therapy of the subject method fora time period of at least about 24 weeks and up to about 48 weeks.

Subjects Suitable for Treatment

Any of the above treatment regimens can be administered to individualswho have been diagnosed with an HCV infection. Any of the abovetreatment regimens can be administered to individuals who have failedprevious treatment for HCV infection (“treatment failure patients,”including non-responders and relapsers).

Individuals who have been clinically diagnosed as infected with HCV areof particular interest in many embodiments. Individuals who are infectedwith HCV are identified as having HCV RNA in their blood, and/or havinganti-HCV antibody in their serum. Such individuals include anti-HCVELISA-positive individuals, and individuals with a positive recombinantimmunoblot assay (RIBA). Such individuals may also, but need not, haveelevated serum ALT levels.

Individuals who are clinically diagnosed as infected with HCV includenaïve individuals (e.g., individuals not previously treated for HCV,particularly those who have not previously received IFN-α-based and/orribavirin-based therapy) and individuals who have failed prior treatmentfor HCV (“treatment failure” patients). Treatment failure patientsinclude non-responders (i.e., individuals in whom the HCV titer was notsignificantly or sufficiently reduced by a previous treatment for HCV,e.g., a previous IFN-α monotherapy, a previous IFN-α and ribavirincombination therapy, or a previous pegylated IFN-α and ribavirincombination therapy); and relapsers (i.e., individuals who werepreviously treated for HCV, e.g., who received a previous IFN-αmonotherapy, a previous IFN-α and ribavirin combination therapy, or aprevious pegylated IFN-α and ribavirin combination therapy, whose HCVtiter decreased, and subsequently increased).

In particular embodiments of interest, individuals have an HCV titer ofat least about 10⁵, at least about 5×10⁵, or at least about 10⁶, or atleast about 2×10⁶, genome copies of HCV per milliliter of serum. Thepatient may be infected with any HCV genotype (genotype 1, including 1aand 1b, 2, 3, 4, 6, etc. and subtypes (e.g., 2a, 2b, 3a, etc.)),particularly a difficult to treat genotype such as HCV genotype 1 andparticular HCV subtypes and quasispecies.

Also of interest are HCV-positive individuals (as described above) whoexhibit severe fibrosis or early cirrhosis (non-decompensated,Child's-Pugh class A or less), or more advanced cirrhosis(decompensated, Child's-Pugh class B or C) due to chronic HCV infectionand who are viremic despite prior anti-viral treatment with IFN-α-basedtherapies or who cannot tolerate IFN-α-based therapies, or who have acontraindication to such therapies. In particular embodiments ofinterest, HCV-positive individuals with stage 3 or 4 liver fibrosisaccording to the METAVIR scoring system are suitable for treatment withthe methods described herein. In other embodiments, individuals suitablefor treatment with the methods of the embodiments are patients withdecompensated cirrhosis with clinical manifestations, including patientswith far-advanced liver cirrhosis, including those awaiting livertransplantation. In still other embodiments, individuals suitable fortreatment with the methods described herein include patients with milderdegrees of fibrosis including those with early fibrosis (stages 1 and 2in the METAVIR, Ludwig, and Scheuer scoring systems; or stages 1, 2, or3 in the Ishak scoring system.).

EXAMPLES 1-5

The syntheses of Compounds 1-5 are summarized in Schemes 1 and 2. Thegeneral procedures below describe the reaction conditions. Compounds 1-5may be prepared using an appropriate sequence of the described generalprocedures. For compounds bearing a carboxylic acid group, it isadvantageous to perform the ester hydrolysis as the last step.

GENERAL PROCEDURE FOR THE CLEAVAGE OF TERT-BUTYL CARBAMATES ANDTERT-BUTYL ESTERS: The compound is dissolved in an appropriate amount ofdichloromethane and 2M HCl in ether (3 equiv.) are added. The reactionis stirred at room temperature until the starting material has beenconsumed. Once complete, the reaction is concentrated in vacuo to obtainthe crude product. In most cases, the crude product is pure enough forfurther transformations. If the crude product is not sufficiently pure,it can be purified using normal or reverse phase chromatography.

GENERAL PROCEDURE FOR THE COUPLING OF AMINES WITH CARBOXYLIC ACIDS: Thecompound is dissolved in an appropriate amount of dichloromethane andthe carboxylic acid (1.1 equiv.) added followed by PS-CDI (2 equiv.) andDIEA (10 equiv.). The reaction is shaken at room temperature until thestarting material has been consumed. Once complete, the reaction isfiltered and the filtrate concentrated in vacuo to obtain the crudeproduct. In most cases, the crude product is pure enough for furthertransformations. If the crude product is not sufficiently pure, it canbe purified using normal or reverse phase chromatography.

GENERAL PROCEDURE FOR THE COUPLING OF AMINES WITH CARBOXYLIC ACIDCHLORIDES: The compound is dissolved in an appropriate amount ofdichloromethane and the carboxylic acid chloride (1.1 equiv.) addedfollowed by PS-DMAP (3 equiv.). The reaction is shaken at roomtemperature until the starting material has been consumed. Oncecomplete, the reaction is filtered and the filtrate concentrated invacuo to obtain the crude product. In most cases, the crude product ispure enough for further transformations. If the crude product is notsufficiently pure, it can be purified using normal or reverse phasechromatography.

GENERAL PROCEDURE FOR THE HYDROLYSIS OF METHYL AND BENZYL ESTERS: Thecompound is dissolved in an appropriate amount of methanol and 3N NaOH(2 equiv.) added. The reaction is stirred at room temperature until allstarting material has been consumed. Once complete, the reaction isconcentrated in vacuo to obtain the crude product. The crude product ispurified by reverse phase chromatography. Table 1 provides LCMSinformation (m/z) for each of the Compounds 1-5.

EXAMPLES 6-10

Compounds 6, 8, 9 and 10 are prepared in the manner described in U.S.patent application Ser. No. 11/093,884, filed Mar. 29, 2005, which ishereby incorporated herein by reference in its entirety, including forthe particular purpose of describing methods of making Compounds 6, 8, 9and 10. Compound 7 is prepared in the manner described in U.S.Provisional Patent Application No. 60/702,195, filed Jul. 25, 2005,which is hereby incorporated herein by reference in its entirety,including for the particular purpose of describing methods of makingCompound 7.

EXAMPLES 11-20

The NS3 protease activity of Compounds 1-10 is indicated in Tables 1-2.NS3 protease activity is determined as described in U.S. patentapplication Ser. No. 11/093,884, filed Mar. 29, 2005, which is herebyincorporated herein by reference in its entirety, including for theparticular purpose of describing methods of determining proteaseactivity.

The NS3 helicase activity indicated in Table 2 is determined as follows:NS3 helicase inhibitory activity is assessed by determining the abilityof the compound to inhibit DNA unwinding in an in vitro homogeneoustime-resolved fluorescence quench assay. The helicase substrate (PerkinElmer, TruPoint Helicase Substrate) consists of partiallydouble-stranded DNA, with one oligonucleotide strand labeled with afluorescent europium chelate and the other strand labeled with the QSY™7 quencher. In the presence of helicase and ATP, this DNA is unwound anda large increase in fluorescence is observed. An excess of an unlabeledoligonucleotide (also from Perkin Elmer, TruPoint Helicase CaptureStrand) that is complementary to the quencher strand is included in theassay to prevent reannealing of the europium and QSY-labeled strands.

The assay buffer contains 25 mM MOPS (pH 7.0), 500 μM MgCl₂, and 0.005%(v/v) Triton X-100, with DMSO being present at a final concentration of2% (v/v). Recombinant, purified, full-length NS3 (1-631) protein isincluded in these assays at a final concentration of 2.5 nM. Thecompound is incubated with NS3 protein for 5 minutes in a 384-well whiteProxiplate™ (Perkin Elmer) prior to the addition of TruPoint HelicaseSubstrate (4 nM final concentration), TruPoint Helicase Capture Strand(15 nM final concentration), and ATP (100 μM final concentration). Thefinal reaction volume is 20 μL. Immediately after the addition of thesubstrates and capture strand, the initial rates of the unwindingreactions are determined at room temperature via an Envision (PerkinElmer) plate reader. The rates of reactions containing test compound arecompared to those lacking test compound in order to evaluate compoundpotency. IC₅₀ values are determined using the curve-fitting softwareXLfit (IDBS).

TABLE 1 No. Structure IC₅₀* LCMS (m/z) 1

D 915.3 2

D 759.3 3

D 915.4 4

D 758.6 5

D 744.4 *NS3 protease activity: A: <50 mM; B: <10 mM; C: <1 mM; D: <0.1mM

TABLE 2 No. Structure IC₅₀ (FL)* IC₅₀ (Helicase)* 6

B 7

B A 8

B B 9

A A 10

B B *FL: Full length NS3; A: <50 mM; B: <10 mM; C: <1 mM; D: <0.1 mM

What is claimed is:
 1. A compound selected from the group consisting of:


2. A pharmaceutical composition, comprising: a pharmaceuticallyacceptable excipient; and a pharmaceutically effective amount of thecompound of claim
 1. 3. A method of modulating the activity of ahepatitis C virus, comprising contacting the virus with an effectiveamount of the compound of claim
 1. 4. The method of claim 3, wherein thecompound modulates NS3 protease.
 5. The method of claim 4, wherein thecompound inhibits NS3 protease.
 6. The method of claim 3, wherein thecompound modulates NS3 helicase.
 7. The method of claim 6, wherein thecompound inhibits NS3 helicase.
 8. The method of claim 3, wherein thecompound modulates both NS3 protease and NS3 helicase.
 9. The method ofclaim 8, wherein the compound inhibits both NS3 protease and NS3helicase.
 10. A method of treating a mammal, comprising: identifying amammal infected with a hepatitis C virus; and administering an effectiveamount of the compound of claim 1 to the mammal.
 11. The method of claim10, further comprising identifying the compound as an inhibitor of bothNS3 protease and NS3 helicase.
 12. A method of treating a mammal,comprising: identifying a mammal infected with a hepatitis C virus; andadministering an effective amount of the pharmaceutical composition ofclaim 2 to the mammal.
 13. The method of claim 12, further comprisingidentifying the compound as an inhibitor of both NS3 protease and NS3helicase.